Thermo Scientific

Reagents for Mass Spectrometry 2015 New Product Guide

New tools

for better mass spec results Extended Mass Range Triple Quad Calibration Solution • BSA and 6 Protein Digest Standards • Antibody Biotinylation Kit • Immunoprecipitation Kits • High pH Reversed-Phase Peptide Fractionation Kit • Quantitative Fluorometric or Colorimetric Peptide Assays

Table of Contents

Introduction

Mass Spectrometry Workflow 

2

Triple Quadrupole Calibration Solution, Extended Mass Range

4

BSA Protein Digest Standard

6

6 Protein Digest Standard

7

Pierce Antibody Biotinylation Kit for IP

8

Pierce MS-Compatible Magnetic IP Kits

9

High pH Reversed-Phase Peptide Fractionation Kit

12

Quantitative Colorimetric Peptide Assay

14

Quantitative Fluorometric Peptide Assay

16

Mass Spec Instruments

18

Mass Spec Rewards Program

20

Related Products 

21

Thermo Fisher Scientific offers a complete portfolio of calibration solutions, standards and sample preparation and quantitation reagents designed for better mass spectrometry (MS) analysis. This portfolio has been developed around the context of biology, and the newest products include an improved triple quadrupole instrument calibration solution, standards of varying complexity to represent typical samples, enrichment kits for the immunoprecipitation of targets of varying abundance and a fractionation spin column kit for deeper peptide coverage. In addition, we developed two innovative assays that can be used to quickly and easily standardize peptide samples following protein digestion.

We recognize the need to provide complete solutions and technical support for proteomics research and analytical analysis using MS instrumentation. These new reagents have been validated to assist the biologist and the mass spectrometrist to succeed in their research. Robust, integrated workflows provide consistent results between labs and eliminate time wasted on troubleshooting experimental methods and results. This brochure highlights the newest products for instrument calibration, quality control and sample preparation to help achieve successful MS analysis.

1

Mass Spectrometry Workflow Tools and reagents for sample preparation and quantitation of proteins using mass spectrometry

CELLS

SERUM, PLASMA AND BIOFLUIDS

TISSUES

METABOLIC LABELING

ABUNDANT PROTEIN DEPLETION

• SILAC stable isotope labeling

• Albumin depletion kits • Abundant protein depletion columns

LYSIS AND PROTEIN EXTRACTION • Cell or tissue lysis reagents • Protease inhibitors • Phosphatase inhibitors

PROTEIN LABELING AND/OR ENRICHMENT • Kinase • GTPase • Serine hydrolase • Phosphoprotein • Immunoprecipitation • Cysteine-reactive mass tagging reagents

IN-SOLUTION DIGESTION • MS-grade proteases • In-solution tryptic digestion kit

PROTEIN CLEAN UP AND QUANTITATION • Dialysis cassettes and devices • Desalting columns • Protein assays

GEL SEPARATION • NuPAGE® Gels • Coomassie stains • Silver stain for MS

2

For more information, or to download product instructions, visit thermoscientific.com/pierce

REAGENTS FOR MS INSTRUMENT CALIBRATION, QC AND ANALYSIS

PEPTIDE CLEAN UP

• MS calibration solutions • MS standards • Ion-pairing reagents

• C18 columns, tips, spin tips • Detergent removal columns • Graphite spin columns

• Chromatography solvents and blends • MALDI matrixes

PEPTIDE ENRICHMENT AND/OR FRACTIONATION • Fe-NTA Phosphopeptide enrichment • TiO2 Phosphopeptide enrichment • High pH Reversed-Phase Peptide Fractionation Kit

Mass Spec Sample Prep Kit for Cultured Cells

PEPTIDE OR GLYCAN LABELING • Amine-reactive mass tagging reagents • Glycan labeling mass tagging reagents

PEPTIDE QUANTITATION ASSAYS • Fluorometric Peptide Assay • Colorimetric Peptide Assay

IN-GEL DIGESTION • In-gel tryptic digestion kit

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

3

Thermo Scientific Pierce Triple Quadrupole Calibration Solution, Extended Mass Range Validated, extended mass range calibration solution that can be used in both positive and negative ionization modes for triple quadrupole instruments

The Thermo Scientific™ Pierce™ Triple Quadrupole Calibration Solution, Extended Mass Range is a mixture of 14 highly pure, ionizable components (mass ranges 69 m/z to 2800 m/z) designed for both positive and negative ionization calibration of Thermo Scientific™ Triple Stage Quadrupole instruments. The Pierce Triple Quadrupole Calibration Solution is a ready-to-use formulation that has been optimized for both positive and negative ionization instrument calibration. The extended mass range (69 m/z to 2800 m/z) calibration has been specifically optimized to improve sensitivity and mass accuracy of the Thermo Scientific™ TSQ Quantiva™ and Thermo Scientific™ TSQ Endura™ Series Triple Quadrupole mass spectrometers. Additionally, the Triple Quadrupole Calibration Solution, Extended Mass Range is manufactured at an ISO 9001 facility and each lot is quality controlled with strict specifications. The stable solution is provided in a leak-proof, high-purity PTFE bottle. Highlights • Improved mass accuracy – extended mass range improves mass accuracy of instrumentation • Positive and negative mode calibration – single formulation with extended mass ranges for both calibration modes • Highly validated – high-quality, rigorously tested formulation with lot-specific certificates of analysis • Ready to use – reference standard may be directly loaded into a syringe and injected into the instrument • Stable – store at 2 to 8°C for 18 months

The Pierce Triple Quadrupole Calibration Solution, Extended Mass Range formulation contains 14 compounds that are ionizable in both positive and negative modes for instrument calibration. The formulation has been specifically optimized for use with the Endura and Quantiva mass spectrometers and is also compatible with the Thermo Scientific™ TSQ Quantum™, Thermo Scientific™ TSQ Discovery™, Thermo Scientific™ TSQ Quantum Ultra™, Thermo Scientific™ TSQ Quantum Access™, and Thermo Scientific™ TSQ Vantage™ Series instruments using the list of recommended masses. • Positive mode (m/z): 69.0447, 142.1590, 322.0481, 622.0289, 922.0098 • Negative mode (m/z): 59.0128, 112.9845, 301.9970, 601.9779, 1033.9870

For the calibration procedure use: TSQ Series Getting Started Guide (70111-97152)

4

Table 1. Accurate mass and specifications of Thermo Scientific Pierce Triple Quadrupole Calibration Solution, Extended Mass Range. Components

Accurate Mass (m/z)

Specification (neat)

*LC-MS grade acetonitrile (92% v/v)

–

Purity 99.9%

*LC-MS grade water (4% v/v)

–

LC-MS Grade

–

Purity 99.9%

*LC-MS grade 2-propanol (4% v/v) LC-MS grade acetic acid

59.0139b(-)

Purity 99.7%

Imidazole

69.0447

Purity 99.0%

Triethylamine

102.1277a(+)

Purity 99.5%

Trifluoroacetic acid (TFA)

112.9856b(-)

Purity 99.5%

Tetramethylpiperidine

a(+)

142.1590

Purity 97.5%

1,8-Bis(dimethylamino) naphthalene

215.1543a(+)

Purity 98.5%

2,4,6-Tris(trifluoromethyl)-1,3,5-triazine

301.9981

Purity > 97.0%

Hexamethoxyphosphazene

322.0481a(+)

Purity 99.0%

2,4,6-Tris(heptafluoropropyl)-1,3,5-triazine

c(-)

601.9779

Purity 95.0%

Hexakis(2,2-difluoroethoxy) phosphazene

622.0290a(+)

Purity 97.0%

Hexakis(2,2,3,3-tetrafluoropropoxy) phosphazene

922.0098 , 1033.9881d(-)

Purity 95.0%

Hexakis(1h,1h,5h-octafluoropentoxy) phosphazene

1521.9715 , 1633.9498d(-)

Purity 95.0%

Hexakis(1h,1h,7h-perfluoroheptoxy) phosphazene

2121.9331a(+), 2233.9115d(-)

Purity 95.0%

Hexakis(1h,1h,9h-perfluorononyloxy) phosphazene

2721.8948a(+), 2833.8731d(-)

Purity 95.0%

a(+)

c(-)

a(+)

a(+)

[M+H] [M-H]c [M+OH]d [M+TFA]*Combined concentration of all other components listed is less than 1% w/v a

+

b

For more information, or to download product instructions, visit thermoscientific.com/pierce

Positive Ion Mode (50-400 m/z)

69.00

100 90 80 70 60 50 40 30 20 10 0

100 90 80 70 60 50 40 30 20 10 0

322.11

142.11 102.11 83.00 215.22

84.00

113.00

100 90 80 70 60 50 40 30 20 10 0

Negative Ion Mode (50-400 m/z) 302.11

69.00

60

80

100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 m/z

100 90 80 70 60 50 40 30 20 10 0 400

50-400 m/z

622.22

922.11 Positive Ion Mode (400-3000 m/z)

1522.11

2122.11

2722.00

Negative Ion Mode (400-3000m/z) 2234.00 602.11

2834.00 1034.11

600

800

1634.11

1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 m/z

400-3000 m/z

100 69.00 90 322.11 80 142.11 70 60 50 40 215.22 30 20 10 0

Positive Ion Mode (50-3000 m/z)

622.22

922.11 1522.11

2122.11

2722.00

2234.00 100 Negative Ion Mode (50-3000 m/z) 90 80 113.00 602.11 70 2834.00 60 302.11 1634.11 50 1034.11 40 30 20 10 0 200 400 600 800 1000 1200 1400 1600 1800 2000 2200 2400 2600 2800 3000 m/z

Figure 1. Calibration solution components and sample chromatograms in positive and negative mode.

Ordering Information Product # Description Pkg. Size 88340 Pierce Triple Quadrupole Calibration Solution, 10mL Extended Mass Range

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

5

Thermo Scientific Pierce BSA Protein Digest Standard, LC-MS Grade High-quality, validated BSA protein digest standard for LC-MS applications

The Thermo Scientific™ Pierce™ BSA Protein Digest Standard, LC-MS Grade is a validated protein digest optimized for use as a quality control sample for liquid chromatography (LC) and MS analysis of proteomic samples. The Pierce BSA Protein Digest Standard is produced using high-quality bovine serum albumin (BSA) and MS-grade trypsin. The BSA is fully reduced, alkylated (iodoacetamide) and desalted (C18) after trypsin digestion to provide a quality LC/MS-grade standard that is free of intact protein. The peptide digest provides excellent sequence coverage with minimal overalkylation and missed cleavages. This digest has been robustly tested for peptide quality, digestion efficiency and lot-to-lot uniformity and is suitable for LC and LC/MS applications.

The Pierce BSA Digest Standard is a lyophilized tryptic peptide mixture that can be used as a quality control standard for liquid chromatography (LC) separation, MS method development and MS performance benchmarking. The digest is specifically formulated for LC/MS experiments and does not contain salts or detergents. Using the digest standard routinely before analysis of samples with similar complexity makes it possible to monitor and normalize LC/MS performance between samples and over time. Moreover, unlike other commercially available protein digests for MS, the Pierce BSA Protein Digest Standard must meet stringent quality testing specifications including peptide quality, digestion efficiency and lot-to-lot digest uniformity.

RT: 0.00-60.00

22.94 653.36

100 90

28.22 582.32

80

Relative Abundance

Highlights • Positive control sample – BSA Protein digest optimized and validated as a quality control standard for MS applications • Excellent sequence coverage – greater than 70% sequence coverage • Validated peptide quality – digestion procedure optimized for minimal missed cleavages and overalkylation • Rigorously tested – high-quality, consistent BSA protein digest guaranteed and documented via lot-specific certificates of analysis • Stable – provided in a stable lyophilized format

70

19.17 722.32 18.78 417.21

60 50

30.06 682.68

40

13.21 488.53

30

10

35.71 700.35

30.74 879.17

8.13 409.72

20

0

24.19 464.25

39.28 784.38

7.30 12.21 545.34 407.21 0

5

10

38.64 997.59

15

20

25

30

35

40

49.78 51.52 430.89 469.18 45

50

60

Figure 1. Thermo Scientific Pierce BSA Digest Standard base peak chromatogram. Base peak chromatogram of 1pmol Pierce BSA Protein Digest Standard separated using a Thermo Scientific™ Acclaim™ PepMap™100 3µm x 75µm x 15cm column (Product # 160321) with a 2-35% gradient (A:0.1% FA in water, B: 0.1% FA in 100% acetonitrile) at 300nl/min for 60 minutes and detected on a Thermo Scientific™ LTQ Orbitrap™ XL mass spectrometer.

Ordering Information Product # Description 88341 Pierce BSA Protein Digest Standard, LC-MS Grade

6

55

Time (min)

For more information, or to download product instructions, visit thermoscientific.com/pierce

Pkg. Size 1nmol

Thermo Scientific Pierce 6 Protein Digest Standard, Equimolar, LC-MS Grade High-quality, validated six protein digest mixture for the standardization of LC-MS applications

The Thermo Scientific™ Pierce™ 6 Protein Digest Standard, Equimolar, LC-MS Grade is a validated protein digest optimized for use as a quality control sample for LC and MS analysis of proteomic samples. The Pierce 6 Protein Digest Standard contains an equimolar mixture of highly pure bovine cytochrome c, lysozyme, alcohol dehydrogenase, bovine serum albumin, apo-transferrin and β-galactosidase. The proteins are reduced, alkylated (iodoacetic acid), digested (MS-grade trypsin) and desalted (C18) after trypsin digestion to provide a quality LC/MS-grade standard free of intact protein. The peptide digest provides excellent sequence coverage with minimal overalkylation and missed cleavages. This digest has been robustly tested for peptide quality, digestion efficiency and lot-to-lot uniformity and is suitable for LC and LC-MS applications.

The Pierce 6 Protein Digest has been optimized as a medium complexity quality control standard for LC-MS applications. It has been optimized to have maximal sequence coverage with minimal missed cleavages and overalkylation. The digest is specifically formulated for LC/MS experiments and does not contain salts or detergents. Using the digest standard routinely before analysis of samples with similar complexity makes it possible to monitor and normalize LC/MS performance between samples and over time. Moreover, unlike other commercially available protein digests for MS, the Pierce 6 Protein Digest Standard must meet stringent quality testing specifications including peptide quality, digestion efficiency and lot-to-lot digest uniformity.

RT: 0.00-120.00

54.06 567.30

100 90

50.49 401.25

80

Relative Abundance

Highlights • Positive control sample – 6 Protein optimized and validated as a quality control standard for MS applications • Excellent sequence coverage – greater than 85% sequence coverage • Validated peptide quality – digestion procedure optimized for minimal missed cleavages and overalkylation • Rigorously tested – high-quality, consistent BSA protein digest guaranteed and documented via lot-specific certificates of analysis • Stable – provided in a stable lyophilized format

70

49.34 418.22

32.39 417.21

60 22.07 407.23

50 40

8.28 550.79

30

16.26 437.71

20

63.28 736.88

79.38 593.36

44.41 653.36

69.36 1043.82 91.90 697.87

23.41 489.19

91.10 784.37

10 0

93.80 108.89 863.41 445.12 0

20

40

60

Time (min)

80

100

120

Figure 1. Thermo Scientific Pierce 6 Protein Digest Standard Base Peak Chromatogram. Base peak chromatogram of 500 fmol Pierce 6 Protein Digest Standard separated using an Acclaim PepMap100 3µm x 75µm x 15cm column (Product # 160321) with a 2-35% gradient (A:0.1% FA in water, B: 0.1% FA in 100% acetonitrile) at 300nl/min for 120 minutes and detected on an LTQ Orbitrap XL mass spectrometer.

Ordering Information Product # Description 88342 Pierce 6 Protein Digest Standard, Equimolar, LC-MS Grade

Pkg. Size 1pmol

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

7

Thermo Scientific Pierce Antibody Biotinylation Kit for IP Optimized antibody biotinylation kits for IP and co-IP applications

The Thermo Scientific™ Pierce™ Antibody Biotinylation Kit for IP provides biotinylation reagents designed specifically for the labeling of primary antibodies used in immunoprecipitation (IP) applications. The Pierce™ Antibody Biotinylation Kit reagents have been optimized and validated to biotinylate antibodies for IP and co-immunoprecipitation (co-IP) reactions. Determining the optimal number of biotins to attach to the target molecule is one of the major challenges of biotinylation. For IP and co-IP applications, too many biotins result in reduced affinity for the target antigen, while too few biotins result in antibody leaching upon elution of the target antigen. The biotin labeling procedure in the Pierce™ Antibody Biotinylation Kit for IP has been developed to address this challenge. Highlights • Optimized – reagents and protocols developed for efficient antibody biotinylation for IP applications • Easy to use – kit contains all reagents and accessories to label and clean-up 50-200µg of antibody • Enhanced solubility – pegylation improves the solubility of the biotinylated antibody and reduces aggregation • Improved binding – longer spacer arm (29 angstroms) on biotin minimizes steric hindrance when binding to avidin molecules

The kit contains sufficient reagents to label 50-200µg of antibody in 100µL reaction volumes for eight samples. The NHS-PEG4-Biotin labeling reagent contains an amine-reactive N-hydroxysuccinimide ester (NHS) group and a water-soluble PEG4 spacer for optimal labeling and is provided in easy-to-use, single-use microtubes. Both the labeling efficiency of the biotinylation reagent and binding affinity of the labeled antibody have been validated using mouse monoclonal (IgG1, IgG2), rabbit polyclonal and rabbit monoclonal antibodies. Thermo Scientific™ Zeba™ Desalting Spin Columns are provided for easy and efficient removal of salts and excess biotin.

Load

MS Bead Elution Boil

Load

MS Bead Elution Boil

PP2A

β-catenin

PTEN

HDAC1 P13K

EGFR

pan Akt

Stat1

Figure 2. The Thermo Scientific Pierce Antibody Biotinylation Kit for IP allows effective target capture and elution. Antibodies were labeled with the kit and used in the Pierce MS-Compatible Magnetic IP Kit (Streptavidin) to IP target proteins from cell lysates. The elutions were analyzed by Western blot.

7 6

Biotins / IgG

5

Ordering Information

4 3

Product # Description 90407 Pierce Antibody Biotinylation Kit for IP

2

Kit contents: EZ-Link™ NHS-PEG4-Biotin Zeba Spin Desalting Columns (7K MWCO), 0.5mL 20X Phosphate Buffered Saline

1 0

Mouse IgG1

Mouse IgG2

Rabbit Mono

Rabbit Poly

Figure 1. The Thermo Scientific Pierce Antibody Biotinylation Kit for IP enables effective labeling of multiple antibody types. The kit was used to label 17 different specific antibodies including mouse IgG1, mouse IgG2, rabbit monoclonal, and rabbit polyclonal. Biotinylation with this kit resulted in three to seven biotins per IgG molecule as determined by the Thermo Scientific™ Pierce™ Fluorescent Biotin Quantitation Kit.

8

90410 90411

For more information, or to download product instructions, visit thermoscientific.com/pierce

Low Protein Binding Microcentrifuge Tubes, 1.5mL Low Protein Binding Microcentrifuge Tubes, 1.5mL

Pkg. Size 8 reactions 8 x 0.5mg 8 columns 15mL

250 tubes 10 x 250 tubes

Thermo Scientific Pierce MS-Compatible Magnetic IP Kits Validated kits for the efficient and reproducible enrichment of target antigens for LC-MS analysis

The Thermo Scientific™ Pierce™ MS-Compatible Magnetic IP Kits provide MS-friendly reagents and optimized protocol to enable highly effective and efficient IP and co-IP of target antigens upstream of LC-MS analysis. In addition, low protein binding microcentrifuge tubes are supplied separately to minimize loss during the sample processing. Highlights • MS-compatible – directly compatible with in-solution peptide digestion • Flexible – different IP strategies are available to utilize either native or biotinylated antibodies • Sensitive – kits have been demonstrated to successfully enrich for low abundant proteins • Low background – buffers optimized to minimize enrichment of background proteins • Robust – procedure and reagents have been robustly tested with numerous targets to ensure consistent enrichment of low abundant proteins

The Pierce MS-Compatible Magnetic IP Kits contain either high-quality Thermo Scientific™ Pierce™ Streptavidin or Protein A/G magnetic beads. The Protein A/G Magnetic Beads provide wider flexibility of antibody capture than either Protein A or G alone. The optimized components of each kit have been formulated to be compatible with downstream LC-MS analysis. After the immunoprecipitation procedure, the target-enriched elution fraction is ready for in-solution tryptic digestion, without the need for gel purification, detergent removal, or desalting. These kits have been rigorously validated using numerous target antigens with varying expression levels, including targets previously undetected without enrichment or by Western blotting. Additionally, the reagents and procedures have been validated using both manual and automated magnetic separation.

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

9

Thermo Scientific Pierce MS-Compatible Magnetic IP Kits (cont.)

Load

MS Bead Elution Boil

Load

MS Bead Elution Boil

PP2A (72%)

β-catenin (140%)

PTEN (90%)

HDAC1 (140%) P13K (247%)

EGFR (80%)

pan Akt (97%)

Stat1 (88%)

Figure 1. The Thermo Scientific Pierce MS-Compatible Magnetic Kit (Streptavidin) allows effective target capture and elution. Antibodies were labeled with the Pierce Antibody Biotinylation Kit for IP and used in the Pierce MS-Compatible Magnetic IP Kit (Streptavidin) to IP target proteins from cell lysates. The elutions were analyzed by Western blot. Percentages beneath target indicate elution efficiency compared to bead boil.

Table 1. Endogenous cellular targets identified with and without enrichment. The Pierce MS-Compatible Magnetic IP Kit (Streptavidin) was used to enrich 17 targets. Antibodies were labeled using the Pierce Antibody Biotinylation Kit for IP. Targets are generally grouped by their relative cellular abundance (high, medium, and low) and may be cell line dependent. Western blots were performed with the IP elutions. Western blot signal intensity is shown as low (+) to high (+++). MS analysis on native, unenriched lysates is indicated by a “yes” when at least two unique peptides were identified for the particular target. MS analysis (enriched) is denoted by a “+++” indicating a higher fold enrichment compared to the MS analysis on native lysate.

Co-IP Proteins

CTNNB1

CTNNA1, CDH2, CDH11, APC, ARVCF, PKP4

EGFR

PRKDC, PFKP, SL C3A2, RPN1

PI3KCA

PIK3R2, PIK3R1

CBP

PSMC5, ACTA2, DDX5

MS analysis (native)

MS analysis (enriched)

+++

yes

++

AKT

VIM, HSPA8, TUBA1A

+++

yes

++

SMAD4

EEF1A1, SQSTM1

+++

yes

++

+

no

+++

PTEN

+++

yes

++

EGFR

++

yes

+++

AKT2

++

yes

+++

AKT1

+

no

+++

CTNNB1

++

no

+++

+

no

+++

+

yes

++

ERBB2

+

no

+++

TP53

+

no

+++

CDH2

-

no

+++

LKB1

ND

no

+++

NOTCH1

ND

no

+++

NOTCH2

ND

yes

+++

PP2A HDAC1 STAT1 CBP

High Medium

PI3K SMAD4

10

IP Target

Western blot (IP)

Target

Cellular abundance

Table 2. List of co-immunoprecipitated proteins. The Pierce MS-Compatible Magnetic IP Kit (Streptavidin) showed effective co-IP of interacting proteins for CTNNB1, EGFR, AKT, PI3KCA, CBP, and SMAD4 targets. These are known protein interactions reported in previous studies.

Low

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MS MS Elution Elution Bead Load 1 2 Boil

MS MS Elution Elution Bead Load 1 2 Boil

HDAC1 (154%)

pan Akt (129%)

PP2A (87%)

β-catenin (112%) Stat1 (108%)

PTEN (719%)

EGFR (122%)

Figure 2. The Thermo Scientific Pierce MS-Compatible Magnetic Kit (Protein A/G) allows effective target capture and elution. Antibodies were labeled with the kit and used in the Pierce MS-Compatible Magnetic IP Kit (Protein A/G) to IP target proteins from cell lysates. The elutions were analyzed by Western blot. Percentages beneath target indicate elution efficiency compared to bead boil.

Table 3. Endogenous cellular targets identified with and without enrichment. The Pierce MS-Compatible Magnetic IP Kit (Protein A/G) was used to enrich 20 targets. Targets are generally grouped by their relative cellular abundance (high, medium, and low) and may be cell line dependent. Western blots were performed with the IP elutions. Western blot signal intensity is shown as low (+) to high (+++). MS analysis on native, unenriched lysates is indicated by a “yes” when at least two unique peptides were identified for the particular target. MS analysis (enriched) is denoted by a “+++” indicating a higher fold enrichment compared to the MS analysis on native lysate. Target PP2A HDAC1 STAT1

Cellular abundance High

Western blot (IP)

MS analysis (native)

MS analysis (enriched)

PI3KCA

PIK3R2, PIK3R1 PTBP1, C14orf166

+++

yes

++

AKT1

AKT2, ACTB

ARAF

YWHAG, STK25

no

+++

PTEN

+++

yes

++

EGFR

++

yes

+++

AKT2

++

yes

+++

AKT1

+

no

+++

CTNNB1

++

no

+++

PI3K

+

no

+++

SMAD4

+

yes

++

+

no

+++

KRAS

TUBB, TUBA1A, HSPA1A

++

+

Low

CTNNA1, CDH11, CDH2, CTNND1

EGFR

yes

++

ERBB2

Co-IP Proteins

CTNNB1

+++

yes

CBP

IP Target

NOTCH1

+++

Medium

Table 4. List of co-immunoprecipitated proteins. The Pierce MS-Compatible Magnetic IP Kit (Protein A/G) showed effective co-IP of interacting proteins for CTNNB1, EGFR, PI3KCA, NOTCH1, AKT1, and ARAF targets. These are known protein interactions reported in previous studies.

+

no

+++

TP53

ND

no

+++

CDH2

ND

no

+++

ARAF

ND

yes

+++

BRAF

ND

no

+++

LOK

ND

no

+++

NOTCH1

ND

no

+++

NOTCH2

ND

yes

+++

Ordering Information Product # Description 90408 Pierce MS-Compatible Magnetic IP Kit (Streptavidin)

Pkg. Size

Kit contents: Streptavidin Magnetic Beads IP-MS Cell Lysis Buffer IP-MS Wash Buffer A IP-MS Wash Buffer B IP-MS Elution Buffer

1mL 100mL 75mL 40mL 6mL

90409

Pierce MS-Compatible Magnetic IP Kit (Protein A/G)

40 reactions

Kit contents: Streptavidin Magnetic Beads Protein A/G Magnetic Beads IP-MS Cell Lysis Buffer IP-MS Wash Buffer A IP-MS Wash Buffer B IP-MS Elution Buffer MgCl2 (1M)

40 reactions

1mL 1mL 100mL 75mL 40mL 6mL 0.75mL

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

11

Thermo Scientific Pierce High pH Reversed-Phase Peptide Fractionation Kit Easy-to-use peptide fractionation kit that reduces sample complexity and increases protein identification

The Thermo Scientific™ Pierce™ High pH ReversedPhase Peptide Fractionation Kit increases protein identification from LC-MS analysis through orthogonal peptide fractionation of complex peptide samples. Highlights • Easy use – resin provided in single-use spin column format • Improved protein identifications – protein identifications increased by ≥ 50% when compared to unfractionated samples • Reproducible – elution profiles and fractional resolution vary by less than 20% • Optimized – robust procedure for maximal protein identification and peptide recovery while minimizing fractional overlap • Compatible – reagents have been validated with a variety of complex samples, including Thermo Scientific™ TMT™ Reagent-labeled peptides

To enable deep proteome sequencing, it is often necessary to reduce the sample complexity by fractionation in an orthogonal dimension prior to LC/MS analysis. The Pierce High pH Reversed-Phase Peptide Fractionation Kit uses high pH reversed-phase chromatography to separate peptides by hydrophobicity and provides excellent orthogonality to low pH reversedphase LC-MS gradients. The kit is designed to improve protein identification through the use of a proprietary reversed-phase resin in an easy-to-use spin column format with a high pH fractionation protocol. In contrast to strong cation exchange (SCX) fractionation, the high pH reversed-phase fractions do not require an additional desalting step before LC/MS analysis.

Column conditioning

Sample loading Conditioned spin column

(1) Acetonitrile (×2) (2) 0.1% TFA (×2)

Centrifuge at 5,000g (2 minutes)

Centrifuge at 3,000g (2 minutes)

Flowthrough fraction Waste

Fractionation and elution

The Pierce High pH Reversed-phase Peptide Fractionation Kit includes a high pH solution (0.1% triethylamine) and 12 spin columns containing pH-resistant, reversed-phase resin. Each reversed-phase fractionation spin column enables fractionation of 10-100µg of peptide sample using a microcentrifuge. Native, phosphorylated, TMT Reagent-labeled and other complex peptide mixture samples can be fractionated using the kit. Combining the search results generated by the individual fractions improves protein sequence coverage and increases number of identified proteins relative to unfractionated samples.

12

Digest sample

Wash

Elution solution 1

Water

Centrifuge at 3,000g (2 minutes)

Centrifuge at 3,000g (2 minutes)

Peptide elution with step gradient

1

2

3 4 5 6 Sample fractions

7

8

Wash fraction

Figure 1. Procedure summary. Peptides are bound to the hydrophobic resin under aqueous conditions and desalted by washing the column with water by low-speed centrifugation. A step gradient of increasing acetonitrile concentrations in a volatile high pH elution buffer is then applied to the columns to elute bound peptides into eight different fractions collected by centrifugation. Each fraction is then dried in a vacuum centrifuge and stored until analysis by MS. During LC-MS analysis, peptides in each high pH fraction are further separated using a low pH gradient, thus reducing the overall sample complexity and improving the ability to identify low-abundant peptides.

For more information, or to download product instructions, visit thermoscientific.com/pierce

55,000 50,000

40,000

A.

35,000

45,000 40,000 35,000 30,000 25,000 20,000 15,000 10,000 5,000 0

Unique Peptide IDs

30,000 25,000 20,000 15,000 10,000

.0 Fr ac Tot tio al na tio n

Fr ac ned tio na tio n

Co

m bi

50

25

.5 22

20

.5

15

17

.5

10

No

Acetonitrile (%)

12

.0

0

50

.5

20

.0

17

.5

15

12

5

.0 10

7.

5.

Flo

wt hr ou

0

5,000

gh W as h

Unique Peptide IDs

A.

No

Acetonitrile (%)

7500

n

d

tio

na

tio

Co

No

Fr

Acetonitrile (%)

ac

No

m

bi

ne

50

25

.5

20

22

.5

0 5.

ac Fr

Acetonitrile (%)

15

n

na

tio

.0

al

To t

tio

50

.0

20

.5

.0

17

15

.5

12

5

.0 10

7.

0

h

5.

W as

Flo

wt

hr

ou

gh

0

17

2500

5,500 5,000 4,500 4,000 3,500 3,000 2,500 2,000 1,500 1,000 500 0

.5

5000 Protein Group IDs

Protein Group IDs

B.

12

B.

C.

100 100 Peptides (%) in Overlapping Fractions

Peptides (%) in Overlapping Fractions

C. 80 60 40 20 0 1

2

3

4

5

6

7

8

60 40 20 0

Number of Fractions

Figure 2. High pH reversed-phase fractionation profile of 100µg HeLa cell lysate tryptic digest. Unique peptides (A) and protein groups (B) identified in each elution fraction compared to the total number of identifications from a combined search of all elution fractions and a single injection of unfractionated sample. Over 100% more unique peptides and over 50% more protein groups are identified in the sample upon high pH reversed-phase fractionation compared to analysis of an no fractionation sample. Excellent fractional resolution is attained (C), with only ~30% fractional overlap. The analysis, performed using triplicate sample sets, shows exceptional reproducibility, as indicated by the very narrow error bars.

80

1

2

3

4

5

6

7

8

Number of Fractions

Figure 3. High pH reversed-phase fractionation profile of 100µg HeLa cell lysate tryptic digest labeled with Thermo Scientific TMT Reagent. Unique peptides (A) and protein groups (B) identified in each elution fraction compared to the total number of identifications from a combined search of all elution fractions and a single injection of unfractionated sample. Over 100% more unique peptides and over 50% more protein groups are identified in the sample upon high pH reversed-phase fractionation compared to analysis of an no fractionation sample. Excellent fractional resolution is attained (C), with only ~30% fractional overlap. The analysis, performed using triplicate sample sets, shows exceptional reproducibility, as indicated by the very narrow error bars.

Ordering Information Product # Description 84868 Pierce High pH Reversed-Phase Peptide Fractionation Kit

Pkg. Size

Kit contents: High pH Reversed-Phase Spin Columns Triethylamine (0.1%)

12 columns 100mL

88379

Low Protein Binding Microcentrifuge Tubes, 2.0mL Low Protein Binding Microcentrifuge Tubes, 2.0mL

250 tubes

88380

Kit

10 x 250 tubes

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

13

Thermo Scientific Pierce Quantitative Colorimetric Peptide Assay Novel colorimetric assay designed specifically for simple, sensitive peptide quantitation

The Thermo Scientific™ Pierce™ Quantitative Colorimetric Peptide Assay is an easy-to-use, colorimetric microplate assay designed specifically for the quantitation of peptide mixtures. Highlights • Sensitive – accurately detect as low as 25µg/mL of peptide mixtures • Robust – assay performance rigorously tested using peptide digest mixtures • Validated standard – kit includes a validated peptide digest standard for improved reproducibility of quantitation • Compatible – can be used directly with most MS sample preparation reagents and TMT Reagent-labeled peptides (Table 1) • Convenient – easy mix-and read-format; colorimetric signal is stable and may be read in 15 minutes

The Pierce Quantitative Colorimetric Peptide Assay provides modified BCA reagents for the reduction of Cu+2 to Cu+1 and a proprietary chelator optimized for the quantitation of peptide mixtures. In this reaction, the copper is first reduced by the amide backbone of peptides under alkaline conditions (Biuret reaction), followed by the proprietary chelator coupling with the reduced copper to form a bright red complex with absorbance at 480nm. The signal produced from this reaction is three- to four-fold more sensitive than the Thermo Scientific™ Pierce™ Micro BCA Protein Assay for peptide analysis. The kit also contains a high-quality peptide digest reference standard to generate linear standard curves and calibration controls. This colorimetric peptide assay requires a small amount of sample (20µL) and has a working peptide concentration range of 25-1000μg/mL. The assay’s increased sensitivity, low sample assay volume and included reference standard are essential for accurate and robust measurement of peptide digest samples, especially for MS applications.

14

Table 1. Compatibility of Thermo Scientific Pierce Quantitative Colorimetric Peptide Assay with commonly used reagents in MS. Substance

Compatible Concentration

Acetone

50%

Acetonitrile

50%

Ammonium bicarbonate

50mM

For more information, or to download product instructions, visit thermoscientific.com/pierce

DMSO

50%

DTT (dithiothreitol)

Not compatible

EDTA

5mM

Formic acid

0.5%

Guanidine

0.25M

Iodoacetamide

1M

Methanol

25%

SDS

1%

Sodium azide

1%

TCEP

Not compatible

TEA acetate

5mM

TEA bicarbonate

5mM

Trifluoroacetic acid

0.5%

Tris

100mM

Urea

1M

A. 2.0

Absorbance (480/562nm)

1.5

Peptide Assay r2=0.996

1.0

Micro BCA Assay r2=0.998

0.5

0 0

100

200

300

400

500

600

Concentration (μg/mL) LOQ (Peptide Assay) = 30μg/mL LOQ (Micro BCA) = 107μg/mL

B. 350 Colorimetric Concentration (μg/mL)

300

Fluorometric

250 200 150 100 50

a A5 49

He L

3T 3 HC T1 16 HE K2 93

31 A4

90 LN Ca p

5

IM R

a

14

DV

He L

pG 2

He

A5

49

0

Cell Lysate

Figure 1. Sensitivity of the Thermo Scientific Pierce Quantitative Colorimetric Peptide Assay. Panel A: Sensitivity of Pierce Quantitative Colorimetric Peptide Assay compared to the Pierce Micro BCA Assay using BSA Digest. Panel B: Quantitation comparison between Pierce Quantitative Peptide Assays using 12 different cultured mammalian cell lysates.

Ordering Information Product # Description Pkg. Size 23275 Pierce Quantitative Colorimetric Peptide Assay 500 assays

23295

Kit contents: Colorimetric Peptide Assay Reagent A Colorimetric Peptide Assay Reagent B Colorimetric Peptide Assay Reagent C Peptide Digest Assay Standard (1mg/mL)

50mL 2 x 25mL 2mL 1.5mL

Peptide Digest Assay Standard

1.5mL

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

15

Thermo Scientific Pierce Quantitative Fluorometric Peptide Assay Novel fluorometric assay designed specifically for simple, sensitive peptide quantitation

The Thermo Scientific™ Pierce™ Quantitative Fluorometric Peptide Assay is a sensitive, mix-and-read fluorescent microplate assay for the quantitative measurement of peptides and peptide mixtures. Highlights • Sensitive – accurately detect as low as 5µg/mL of peptide mixtures • Robust – assay performance rigorously tested using peptide digest mixtures • Validated standard – kit includes a validated peptide digest standard for improved reproducibility of quantitation • Compatible – can be used directly with most MS sample preparation reagents (Table 1) • Convenient – easy mix-and-read format; fluorescent signal is stable and may be read in 5 minutes

The Pierce Quantitative Fluorometric Peptide Assay reagents include peptide assay buffer, fluorescent peptide labeling reagent and a peptide digest assay standard for the quantitative measurement of peptide concentrations. In this assay, peptides are specifically labeled at the amino-terminus using an amine-reactive fluorescent reagent, and the fluorescently labeled peptides are detected using ex390/em475. Because of the labeling mechanism of the fluorescent assay reagent, this assay is suitable for the quantitative measurement of synthetic peptides as well as peptide digest mixtures. The high-quality peptide digest reference standard is included to generate linear standard curves and calibration control for improved accuracy and reproducibility. This sensitive assay requires only 10µL of sample, produces a linear response with increasing peptide concentrations (5-1000μg/mL) and results in a stable final fluorescence that can be detected in as few as five minutes.

16

Table 1. Compatibility of Thermo Scientific Pierce Quantitative Fluorometric Peptide Assay with commonly used reagents in MS. Substance

Compatible Concentration

Acetone

25%

Acetonitrile

50%

Ammonium acetate

100mM

Ammonium bicarbonate

50mM

DMSO

50%

DTT (dithiothreitol)

10mM

EDTA

25mM

Formic acid

0.1%

Guanidine

1M

Iodoacetamide

100mM

Methanol

25%

SDS

1%

Sodium azide

1%

TCEP

10mM

TEA acetate

100mM

TEA bicarbonate

100mM

Trifluoroacetic acid

0.2%

Urea

1M

For more information, or to download product instructions, visit thermoscientific.com/pierce

A.

B.

control 1

175 1000 Measured Concentration (μM)

Fluorescence (A.U.)

150 125 100 75 50 25 0

3 750

4 5 6

500

7 8

250

9 10

0 0

200

400

600

800

0

1000

100

200

300

400

175

Peptide

Sequence

1

SYSMEHFRWG

150

2

RPKPQQFFGLM-NH2

125

3

DRVYIHPFHL

4

RPPGFSPFR

100

5

CYIQNCPLG

75

6

FGGFTGARKSARK

7

GSNKGAIIGLM

50 25 0 0

200

400

600

500

600

800

1000

Concentration (μg/mL)

8

KRPPGFSPFR

9

CYFQNCPRG - NH2

10

INLKALAALAKKIL - NH2

11

DRVYIHPF

12

QKRPSQRSKYL

11 12

Weighed Concentration (μM)

Concentration (μM)

Fluorescence (A.U.)

2

Figure 1. Standard curves for Thermo Scientific Pierce Fluorometric Peptide Assay. Panel A: Pierce Peptide Digest Assay Standard curves using the Pierce Quantitative Fluorometric Peptide Assay. Panel B: Quantitation of individual peptides using the Pierce Quantitative Fluorometric Peptide Assay.

Ordering Information Product # Description 23290 Pierce Quantitative Fluorometric Peptide Assay

Pkg. Size

Fluorometric Peptide Assay Reagent Fluorometric Peptide Assay Buffer Peptide Digest Assay Standard (1mg/mL)

4 x 2.5 mL 50mL 1.5 mL

23295

Peptide Digest Assay Standard

1.5mL

88378

96-well Black Plate

25 pack

500 assays

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

17

Mass Spec Instruments Featured liquid chromatography and mass spectrometry instrumentation and software for proteomics applications

Orbitrap Fusion Tribrid Mass Spectrometer The Thermo Scientific™ Orbitrap Fusion™ mass spectrometer with its revolutionary Tribrid™ architecture provides unprecedented depth of analysis for complex biological challenges. The new 1.1 Instrument Control Software and operating modes bring powerful new features to enable researchers to focus on their science rather than experimental setup.

EASY nLC 1000 Liquid Chromatography The Thermo Scientific™ EASY-nLC™ 1000 nanoflow LC delivers sharper peaks for better resolution and increased protein coverage with accurate and precise solvent flow management. Dual flow sensors placed ahead of the mixer ensure precise gradients for reproducible results. Thermo Scientific™ Dionex™ nanoViper™ fingertight fitting system technology and a large touch screen make the system easy to set up and use.

Q Exactive HF Benchtop Orbitrap Mass Spectrometer The Thermo Scientific™ Q Exactive™ HF mass spectrometer is the next generation benchtop system with an ultra High Field Orbitrap detector, providing speed, productivity and confidence for both qualitative and quantitative omics workflows.

18

For more information, or to download product instructions, visit thermoscientific.com/pierce

TSQ Quantiva Triple Quadrupole Mass Spectrometer The Thermo Scientific™ TSQ Quantiva™ Triple quadrupole mass spectrometer with Active Ion Management (AIM) technology delivers unrivaled sensitivity, speed, and dynamic range for scientists facing the most difficult quantitative challenges now and in the future. This extreme performance is achieved with a reliability and ease of operation that help users spend more time thinking about their analyses and less time worrying about instrument setup and operation.

Integrated Proteomics Software A full suite of software packages are available for your proteomics workflow from discovery through to verification and validation. Products include Thermo Scientific™ Proteome Discoverer™ Software for qualitative and quantitative analysis of proteomics data, Thermo Scientific™ SIEVE™ software for differential analysis, Thermo Scientific™ Pinpoint™ Software for the creation of targeted quantitative assays and specialist software for glycoproteomics. All are designed to take full advantage of the high-quality, high-resolution, accurate-mass data produced by Thermo Scientific™ Orbitrap™-based mass spectrometers.

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

19

Mass Spec Rewards Program and Handbooks Save money and learn more about Thermo Scientific reagents and kits for proteomics applications using mass spectrometry

Through the Thermo Scientific™ Mass Spec Rewards Program any mass spec facility can save 40% on a single order up to $2,500 or the local currency equivalent. Save even more if you purchase one or more Thermo Scientific™ Mass Spectrometers in 2015. To make a purchase through this program, please contact your local sales representative for a list of qualifying products, and a discounted quote on the items selected using the appropriate Promo Code. Reward Levels Customers who

Promo Code

Purchase up to

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MSReward2

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40% off

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Contact your local sales representative with any questions regarding this program. This offer is limited to one order per facility per calendar year and cannot be combined with other offers. This offer is good through December 31, 2015.

Get the NEW Mass Spectrometry Technical Handbooks Download or order our three new technical handbooks to learn how you can optimize your mass spectrometry experiments with Thermo Scientific™ Pierce™ Protein Biology Products.

Protein Quantitation Using Mass Spectrometry

Sample Preparation for Mass Spectrometry

Thermo Scientific Pierce Products for Mass Spectrometry Sample Preparation

* * O

O

O

* * * NH

O

* O

O

*N

N * * H

N O

* *

O O

*

N

O

* O *N

H

Thermo Scientific Pierce Reagents for Quantitative Proteomics

for mass spectrometry

Tools and reagents to extract, digest, enrich and clean up proteins and peptides

20

Protein quantitation tools and specialized reagents for discovery and targeted proteomics Product # 1602351

protein quantitation using mass spectrometry

Specialized reagents for discovery and targeted analysis

Calibration Solutions for Mass Spectrometers

Tools and reagents to extract, digest, enrich and clean up proteins and peptides in one volume

Thermo Scientific Pierce Controls and Standards for Mass Spectrometry

Product # 1602773 sample preparation

N

O

*

O

O

* * NH

O

N N

O O

O

*

O O N * H

Go to piercenet.com/cat/promotional-literature

Three valuable assets for your mass spectrometry experiments:

O

N * H

O

Ensure confidence in instrument performance with Thermo Scientific™ Pierce™ Calibration Solutions and Standards Product # 1602867

calibration tools

for mass spectrometry Ensure confidence in instrument performance with Thermo Scientific Pierce Calibration Solutions and Standards.

For more information, or to download product instructions, visit thermoscientific.com/pierce

Related Products

Calibration Solutions and Standards Pierce™ Peptide Retention Time Calibration Mixture, 0.5pmol/µL Pierce Peptide Retention Time Calibration Mixture, 5pmol/µL

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Abundant Protein Removal

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88320

Albumin Depletion Kit

85160

24-rxn kit

Top 2 Abundant Protein Depletion Spin Columns

85161

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88321

200µL

Top 2 Abundant Protein Depletion Spin Columns

85162

Pierce LTQ ESI Positive Ion Calibration Solution

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Pierce™ LTQ Velos ESI Positive Ion Calibration Solution

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88323

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Top 12 Abundant Protein Depletion Spin Columns

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Pierce™ ESI Negative Ion Calibration Solution

88324

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Pierce™ Triple Quadrupole Calibration Solution

88325

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Protein Digestion – Mass Spec Grade Proteases, Reagents and Kits

Package Product # Size

Reserpine Standard for LC-MS

88326

5 x 1mL

Trypsin Protease, MS Grade

90057

5 x 20µg

Pierce™ HeLa Protein Digest Standard

88328

20µg

Trypsin Protease, MS Grade

90058

5 x 100µg

Pierce HeLa Protein Digest Standard

88329

5 x 20µg

Trypsin Protease, MS Grade

90059

1mg

Digestion Indicator for Mass Spectrometry

84841

10µg

Trypsin Protease, MS Grade, Frozen Liquid

90305

100µg

LysN Protease, MS Grade

90300

20µg

LysN Protease, MS Grade

90301

5 x 20µg

™

50µL

Protein Quantitation Reagents – Amine-Reactive Tandem Mass Tag™ Reagents

Package Product # Size

TMTduplex Isobaric Mass Tagging Kit

90063

15-rxn kit

LysC Protease, MS Grade

90051

20µg

TMTsixplex™ Isobaric Mass Tagging Kit

90064

35-rxn kit

AspN Protease, MS Grade

90053

2µg

TMTduplex Isobaric Label Reagent Set

90065

10-rxn kit

GluC Protease, MS Grade

90054

5 x 10µg

TMTsixplex Isobaric Label Reagent Set

90066

30-rxn kit

Chymotrypsin (TLCK treated) , MS Grade

90056

4 x 25µg

TMTzero™ Label Reagent

90067

5 x 0.8mg

In-Gel Tryptic Digestion Kit

89871

Kit

TMTduplex Isotopic Label Reagent Set

90060

10-rxn kit

In-Solution Tryptic Digestion and Guanidination Kit

89895

Kit

TMTsixplex Isobaric Label Reagent Set

90061

6-rxn kit

Mass Spec Sample Prep Kit for Cultured Cells

84840

20-rxn kit

TMTsixplex Isobaric Label Reagent Set

90062

12-rxn kit

TMTsixplex Isobaric Label Reagent Set

90068

72-rxn kit

Phosphopeptide Enrichment and Clean-Up

Package Product # Size

TMT10plex™ Label Reagent Set

90110

10-rxn kit

Pierce™ Fe-NTA Phosphopeptide Enrichment Kit

88300

30-rxn kit

TMT10plex Label Reagent Set

90111

30-rxn kit

88301

24-rxn kit

TMT10plex Isobaric Mass Tag Labeling Kit

90113

30-rxn kit

TMT10plex Isobaric Label Reagent Set

90406

60-rxn set

88811

96-rxn kit

Protein Quantitation Reagents – Peptides for Targeted Quantitation HeavyPeptide™ AQUA BASIC, custom peptide, up to 15 aa, >95% purity, AAA, dried LightPeptide™ AQUA BASIC, custom peptide, up to 15 aa, >95% purity, AAA, dried HeavyPeptide™ AQUA QuantPro, custom peptide kit, up to 15 aa, >97% purity, AAA ~10-25%, solubilized concentration adjusted LightPeptide™ AQUA QuantPro, custom peptide kit, up to 15 aa, >97% purity, AAA, ~10-25%, solubilized concentration adjusted HeavyPeptide™ AQUA Ultimate, custom peptide kit, up to 15 aa, >97% purity, AAA ~5-10%, concentration adjusted LightPeptide™ AQUA Ultimate, custom peptide kit, up to 15 aa, >97% purity, AAA ~5-10%, concentration adjusted PEPotec™ SRM Peptide, G1, custom crude peptides in 2d-barcoded 96-well library format, 6-25 aa, unmodified PEPotec™ SRM Peptide, G1, heavy labeled, custom crude peptides in 2d-barcoded 96-well library format, 6-25 aa HeavyPeptide IGNIS Prime; up to 20 aa, custom peptide sequences including reporter plus calibration mixture - quantitation kit

Package Product # Size 1x 300206 30nmol 1x 300207 30nmol

Pierce Magnetic TiO2 Phosphopeptide Enrichment Kit 88812

24-rxn kit

Pierce Graphite Spin Columns, 0.5mL

30 columns

™

300204

10 x 1nmol

300205

10 x 1nmol

Pierce™ TiO2 Phosphopeptide Enrichment and Clean-Up Kit Pierce™ Magnetic TiO2 Phosphopeptide Enrichment Kit

™

Peptide Clean-Up Pierce™ Detergent Removal Spin Column, 125µL Pierce Detergent Removal Spin Column, 0.5mL

300215

500200

10 x 1nmol 10 x 1nmol 0.1mg

87778

5 columns

Pierce Detergent Removal Spin Column, 4mL

87779

5 columns

Pierce Detergent Removal Resin

87780

10mL

Pierce™ Detergent Removal Spin Plates

88304

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HiPPR Detergent Removal Spin Column Kit (Resin + Columns)

88305

5mL kit

HiPPR Detergent Removal Spin Columns, 0.1mL

88306

24 columns

HiPPR Detergent Removal 96-well Spin Plates

88307

2 plates

Pierce C18 Spin Columns

89870

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Pierce C18 Tips, 10µL bed

87782

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Pierce C18 Tips, 100µL bed

87784

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Pierce C18 Spin Tips

84850

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0.1mg

300322

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Package Product # Size 25 87776 columns 25 87777 columns

Pierce Detergent Removal Spin Column, 2mL ™

300210

88302

To order, call 800.874.3723 or 815.968.0747. Outside the U.S., contact your local branch office or distributor.

21

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