Deglycosylation Using TFMS
Glycosylation is one of the most common posttranslational modifications of proteins. Glycans attach to the peptide backbone through either amide or glycosidic bonds (N-linked or O-linked) and may be removed either enzymatically or chemically. Enzymatic modes offer mild conditions, but are specific for either N- or O-linked glycans. Chemical methods though are non-specific, but capable of removing the entire glycan complement under appropriate reaction conditions. TFMS is a strong acid, achieves rapid, non-selective deglycosylation without compromise of the protein moiety, permitting subsequent analysis for proteomics and many other downstream analytical applications
Deglycosylation Using TFMS
(Perform in a Fume Hood)
Open TFMS and add toluene to final concentration of 10% (transfer 180mL TFMS to vial and add 20mL toluene) (aliquot remaining TFMS into glass vials – after finish adding mix to samples!)
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