Product Class: Other

O-Glycoprotease (IMPa)
Gray property icon for cloned at NEB recombinant Gray property icon for incubation temperature 37° 95 Heat

Effective 9/27/2022, name modified to O-Glycoprotease (IMPa)

Product Introduction

O-Glycoprotease (IMPa) is a broad specificity protease that cleaves the peptide bonds of a glycoprotein or glycopeptide immediately N-terminal to a serine or threonine residue containing a mucin-type O-linked glycan with or without sialylation.

  • Enables O-glycosylation site mapping and O-glycan structural profiling
  • Efficiently cleaves adjacent to O-glycans with or without sialic acid, no neuraminidase treatment necessary
  • Recombinant enzyme with no detectable non-specific protease contaminating activities
  • Can be used under native or denaturing conditions
  • Glycerol-free for optimal performance in HPLC and mass spectrometry analysis
  • Supplied in solution; optimal activity and stability for up to 24 months
 
Bioz Badge Exists : True
Catalog # Size Concentration
P0761S 200 reactions 1000 units/ml

Product Information

Description

O-Glycoprotease (IMPa), also known as Immunomodulating protease (IMPa), is a broad specificity O-glycoprotease from Pseudomonas aeruginosa.  It recognizes mucin-type O-glycans in glycoproteins or glycopeptides, including branched and sialylated structures, and cleaves the peptide bond immediately N-terminal to a glycosylated serine or threonine residue. It can be used in glycoproteomics workflows to map O-glycosylated residues and structurally profile O-glycans present at each glycosite. O-Glycoprotease (IMPa) has a 6xHis-tag for easy removal from a reaction using nickel affinity resins.

 

Specificity:


Product Source

Cloned from Pseudomonas aeruginosa and expressed in E. coli.
This product is related to the following categories:
Proteases,
Protein Tools,
Glycobiology,
Proteome Analysis,
This product can be used in the following applications:
Glycomics and glycoproteomics,
Protein Digestion,
Glycobiology & Proteomics,
Proteomics,
Glycoprotein Analysis

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit of O-Glycoprotease (IMPa) will cleave > 90% of 2 pmol FAM-labelled O-glycopeptide in a total reaction volume of 20 µl in 2 hours at 37°C in 20mM Tris-HCl, pH 8.0.

Storage Buffer

20 mM Tris-HCl
100 mM NaCl
pH 7.5 @ 25°C

Heat Inactivation

95°C for 10 minutes

Molecular Weight

Theoretical: 97 kDa

Unit Assay Conditions

Two-fold dilutions of O-Glycoprotease (IMPa) are incubated with 2μM of FAM-labelled O-glycopeptide and 20 mM Tris-HCl pH 8.0 in a 20 µl reaction. The reaction mix is incubated at 37°C for 2 hours. Separation of reaction products are determined by CE analysis.

Features

  • O-glycosylation site mapping
  • O-glycan structural determination
  • Enables characterization of simple and complex glycoproteins containing O-linked glycans
  • Efficiently cleaves glycoproteins with or without sialic acid, no neuraminidase treatment necessary
  • 200 reactions (one vial) is sufficient for the cleavage of up to 2 mg of glycoprotein

Product Notes

  1. O-Glycoprotease (IMPa) cleaves a variety of glycoproteins and glycopeptides containing mucin-type O-glycans with or without sialic acid, including Tn-antigen (GalNAc-α-Ser/Thr).
  2. Glycoproteins can be sequentially digested with O-Glycoprotease (IMPa) followed by Trypsin-ultra Mass Spectrometry Grade (NEB #P8101).
  3. The optimal reaction buffer for O-Glycoprotease (IMPa) activity is 20 mM Tris-HCl, pH 8.0. The optimal pH range for O-Glycoprotease (IMPa) is pH 7.0-8.0. Enzyme activity decreases significantly at pH values below 7.0 and above pH 8.0.

FAQs & Troubleshooting

FAQs

  1. Can an O-Glycoprotease (IMPa) reaction be scaled linearly to accommodate digestion of greater than 10 μg of glycoprotein?
  2. Is O-Glycoprotease (IMPa) active in the presence of reducing agents or detergents?
  3. Is O-Glycoprotease (IMPa) active in buffers other than 20 mM Tris-HCl pH 8.0?
  4. Is O-Glycoprotease (IMPa) able to efficiently cleave clustered O-glycosylated sites?
  5. I see minimal cleavage of my glycoprotein after treatment with O-Glycoprotease (IMPa). How can I optimize the reaction?
  6. Is it necessary to first remove sialic acids from the substrate prior to digest with O-Glycoprotease (IMPa)?
  7. Is O-Glycoprotease (IMPa) active on TF-antigen and Tn-antigen?
  8. Does O-Glycoprotease (IMPa) require metal ions for activity?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specification Change Notifications

Effective 9/27/2022, name modified to O-Glycoprotease (IMPa)

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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