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Protein Phosphorylation

Proteoprofile™ Kit

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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