Easily and seemlessly prepare protein samples from stained gels for proteomics analysis by mass spectrometry.

The In-Gel Tryptic Digestion Kit provides a complete set of reagents for performing approximately 150 digestions on colloidal coomassie- or fluorescent dye-stained protein bands. The kit includes modified porcine trypsin, destaining buffers, reduction reagents, alkylation reagents and digestion buffers along with detailed and simple instructions. Each component and step has been optimized and balanced to produce complete, accurate and clean digests using a variety of conditions for dependable proteomics analysis of peptide profiles by mass spectrometry (MS).

Accurate identification and analysis of proteins is a central component of proteomic strategies for studying cellular functions and processes. Mass spectrometers are essential tools for these studies as they provide a high level of sensitivity and mass accuracy. However, this sensitivity and accuracy can be obtained only if proper sample preparation is performed. Two key sample preparation steps are the fragmentation of proteins into peptides and subsequent concentration and clean up of peptides. The In-Gel Tryptic Digestion Kit accomplishes the first of these steps. Thermo Scientific Pierce C-18 Columns accomplish the second step.

Highlights:

• Convenient – includes all necessary reagents for destaining coomassie- or fluorescent dye-stained proteins, reduction and alkylation of cystines, and tryptic digestion
• Robust – the procedure and reagents produce reliable digestions and data generation using a wide range of conditions without optimization
• Accurate – contains highly purified and modified MS-grade trypsin that shows no chymotryptic activity and minimal autolytic activity

Procedure summary for the In-Gel Tryptic Digestion Kit.

Clean MALDI-TOF spectra from samples obtained using the In-Gel Tryptic Digestion Kit. MALDI-TOF MS analysis of bovine serum albumin (BSA) digest. Ten nanograms (150 fmol) of BSA was separated by SDS-PAGE and stained with GelCode Blue Stain Reagent (Product #24592) and then processed with the In-Gel Tryptic Digestion Kit. The resulting digest was treated with Pierce C-18 Spin Columns (Product #89870) then subjected to analysis on a Applied Biosystems VoyagerDE-PRO MALDI-MS in positive ion, linear, delayed-extraction mode. Database searches identified BSA with 47.0% sequence coverage.

Related Products:

GelCode Blue Stain Reagent
Pierce C-18 Spin Columns
Krypton Fluorescent Protein Stain
Pierce Silver Stain for Mass Spectrometry
In-Solution Tryptic Digestion and Guanidination Kit