ProteoProfile™ Enzymatic In-gel N-Deglycosylation Kit, (PP0200)

Most proteins in eurkaryotic cells undergo post-translational modification, quite commonly glycosylation. Unfortunately, this type of modification leads to problems in subsequent protein analysis procedures. Glycopeptides generally do not readily ionize during MS analysis leading to insufficient spectral data. Furthermore, proteolytic digestion of the native glycoprotein is often incomplete due to steric hindrance from the bulky oligosaccharides attached to the protein molecule. Hence removal of the carbohydrate groups from a glycoprotein prior to protein identification is preferred.

Sigma's ProteoProfile™ Enzymatic In-gel N-Deglycosylation Kit is optimized to provide a convenient and reproducible method to N-deglycosylate and tryptically digest protein samples from 1D or 2D polyacrylamide gel slices for subsequent MS or HPLC analysis. The procedure is suitable for Coomassie Blue and Colloidal Coomassie stained gels and may be used with gels silver stained and destained using Sigma's Proteo Silver™ Plus kit ( PROT-SIL2). ProteoProfile Enzymatic In-gel N-Deglycosylation kit includes the enzymes and reagents necessary for N-linked deglycosylation and tryptic digestion. The samples can then be desalted and concentrated for analysis by MALDI-TOF or electrospray MS.

Kit Components     Destaining Solution     Proteomics Grade PNGase F     Proteomics Grade Trypsin     Trypsin Solubilization Reagent     Trypsin Reaction Buffer     Ribonuclease B (positive control)     Peptide Extraction Solution     Biotech Grade Acetonitrile

Features & Benefits

• Provides all components for in-gel deglycosylation AND trypsinization of protein samples
  Conveniently prepares deglycosylated protein samples for analysis by MS or HPLC
• Utilizes PNGase F for the enzymatic removal of N-linked glycans
  Proteins remain intact, unlike the use of chemical deglycosylation which can degrade the protein
• Includes Proteomics Grade PNGase F and Trypsin
  Highly purified enzymes possess no unwanted activities or additives to complicate analysis.
• PNGase F is supplied lyophilized from a low salt buffer
  Allows reconstitution of the enzyme to any concentration needed
• Detailed protocol
  Easy to follow protocol simplifies the process
• Works in solution or with gel slices
  Allows choice of methods

View Protocol Diagram

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