Competitive Advantages

Trust your quantitation!

Multinotch MS3 quantitation is more accurate than other MS2 Methods

The accuracy of Multinotch MS3 quantitation means not missing important expression level changes due to co-isolated interference

Multinotch MS3 quantitation is only available on the Orbitrap Fusion and Orbitrap Fusion Lumos

Orbitrap Fusion Lumos provides highest sensitivity, highest selectivity and lowest detection limit for best quantitation

Sample Preparation

Sample Preparation: Materials

Sample Preparation: Simple Peptide Labeling

Reduced and alkylated trypsin digested proteins

Use Non-Amine Buffer @ pH ~ 8.0 (e.g. TEAB)

Part No. 88328

Description：HeLa Protein Digest Standard Formulation: Lyophilized peptide mixture from a

tryptic digest of HeLa S3 cell lysate Sufficient For: 20 to 100 analyses

•Add 41μL of anhydrous acetonitrile to each tube. Allow the reagent to dissolve for 5 minutes

 with occasional vortexing. Briefly centrifuge the tube to gather the solution.

•Transfer 25-100 uL of the reduced and alkylated protein digest (each condition) to the TMT

 Reagent vial (41 uL). Add sufficient 100 mM TEAB buffer to reach a final volume in vial of 141 uL. Vortex briefly

•Incubate the reaction for 1 hour at room temperature.

•Add 8μL of 5% hydroxylamine to the sample and incubate for 15 minutes to quench the reaction.

•Combine samples in a new microcentrifuge tube at equal amounts and speed vacuum to dryness to

 remove all TEAB

•Aliquot and Store at -80°C.

A More Complete Workflow For Better Coverage

High pH Fractionation Spin Columns

Thermo Poster Note 64606: High pH Reversed-Phase Peptide Fractionation in a Convenient SpinColumn Format; Snovida S. et al

Thermo Poster Note 64604: Quantitative peptide assay for optimized and reproducible sample preparations for mass spectrometry applications; Jiang X. et al

High pH Reversed Phase vs SCX Fractionation