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Protein Digestion
Product Listing
Application Overview
Proteins found in nature vary greatly in size from 5 kDa to greater than 400 kDa. While it is possible to study intact proteins by mass
spectrometry (MS) and the modifications present on these proteins, the most common proteomic approaches currently utilize digestion with
site-specific proteases to generate smaller fragments, peptides, as a first digestion with site-specific proteases to generate smaller
fragments, peptides, as a first step in the analyses (protein digestion). Peptides are easier to characterize and can be separated using
reverse phase supports by high performance liquid chromatography (HPLC) using a C18 column. HPLC-coupled to a Tandem MS is used to
obtain fragmentation data of individual peptides. This digestion of proteins into smaller pieces is typically carried out by proteases such as
trypsin (NEB #P8101) and Endoproteinases GluC (NEB #P8100) and AspN (NEB #P8104).
Featured Products
Endoproteinase GluC
Trypsin-ultra™, Mass Spectrometry Grade
Endoproteinase AspN
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FAQs for Protein Digestion
FAQs
Protocols
Other Tools &
Resources
Legal Information
I have a very low concentration of protein and would prefer not to denature as a separate step with buffer
exchange before digestion. What denaturants can he use in the Trypsin reaction itself?
I am using Trypsin and am wondering about specificity. Does it cut at additional sites when in high
concentration?
How does one do a Trypsin in-gel digest?
Is there a simple way to remove Trypsin after protein cleavage?
Which residues does Endoproteinase AspN cut?
What is the Proteinase K activity in commonly used buffers?
Protocols for Protein Digestion
O-Glycosidase (P0733)
O-Glycosidase Application Note 1 (P0733)
Endo-α-N-Acetylgalactosaminidase Application Note 1
In-gel Digestion Protocol for Endoproteinase LysC (P8109)
Protocol using Trypsin-ultra™, Mass Spectrometry Grade (P8101)
Reaction Protocols for Protein Deglycosylation Mix II (P6044)
Removal of terminal N-acetylglucosamine from the biantennary N-linked sugars of IgG
RNase B Deglycosylation Protocol (P7817)
Trypsin Digestion Protocol using NEB Trypsin-ultra™ and the FASP Kit
Typical GluC Digest Reaction Conditions (P8100)
Typical GluC In-Gel Digest Reaction Conditions (P8100)
Typical Reaction Conditions (P0732)
Typical Reaction Conditions for α2-3 Neuraminidase S (P0743)
Typical Reaction Conditions for β-N-Acetylglucosaminidase S (P0744)
Typical Reaction Conditions for β1-4 Galactosidase S (P0745)
Typical Reaction Conditions for β1-4 Galactosidase (P0730)
Typical Reaction Conditions for Endoproteinase LysC (P8109)
Typical Trypsin and GluC Co-Digest Reaction Conditions (P8100)
Other Tools & Resources
Brochures
Glycobiology Brochure
The Glycobiology brochure provides information on the suite of endo- and exoglycosidases, heparinases
and proteases offered by NEB.
Legal and Disclaimers
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New
England Biolabs, Inc (NEB).
While NEB develops and validates its products for various applications, the use of this product may require
the buyer to obtain additional third party intellectual property rights for certain applications.
For more information about commercial rights, please contact NEB's Global Business Development team at
[email protected].
This product is intended for research purposes only. This product is not intended to be used for therapeutic or
diagnostic purposes in humans or animals.
Related Applications
Glycobiology
Protein Expression & Purification