Product Class: Other

IdeZ Protease (IgG-specific)
Gray property icon for NEBU Gray property icon for cloned at NEB recombinant Gray property icon for incubation temperature 37° Gray property icon for heat inactivation 65°

Product Introduction

IdeZ Protease (IgG-specific) is a recombinant antibody specific protease that recognizes human, sheep, monkey, rabbit, and some murine IgG subclasses, cleaving specifically at a single recognition site below the hinge region, to yield a homogenous pool of F(ab´)2 and Fc fragments. IdeZ Protease more effectively cleaves murine IgG2a than IdeS.

  • Complete fragmentation of antibodies and immunoglobulin fusion proteins in 30 minutes under native conditions
  • Supplied in solution; optimal activity and stability for up to 24 months
  • Combine with PNGase F for complete fragmentation and deglycosylation
  • Recombinant enzyme with no detectable non-specific protease contaminating activities
  • Glycerol-free for optimal performance in HPLC and mass spectrometry analysis
  • ≥95% purity, as determined by SDS-PAGE and intact ESI-MS
Bioz Badge Exists : True
Catalog # Size Concentration
P0770S 4000 units 80000 units/ml

Product Information

Description


 
IdeZ Protease (IgG-specific) is a recombinant antibody specific protease that recognizes human, sheep, monkey, rabbit, and some murine IgG subclasses, specifically cleaving at a single recognition site below the hinge region, yielding a homogenous pool of F(ab´)2 and Fc fragments. IdeZ Protease more effectively cleaves murine IgG2a than IdeS.

Digestion of lgG with IdeZ Protease (lgG-specific), followed by denaturation.

Product Source

Cloned from Streptococcus equi subspecies zooepidemicus and expressed in E. coli. 
This product is related to the following categories:
Proteases,
Proteome Analysis,
This product can be used in the following applications:
Biotherapeutics and Antibody Analysis,
Proteomics

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • NEB # Stored at (°C)
    P0770S -20
    Component Name Component # Stored at (°C) Amount Concentration
    IdeZ Protease (IgG-specific) P0770SVIAL -20 1 x 0.05 ml 80,000 units/ml
    GlycoBuffer 2 B3704SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to cleave > 95% of 1 µg of human IgG, in 15 minutes at 37°C in a total reaction volume of 10 µl.

Reaction Conditions

1X GlycoBuffer 2
Incubate at 37°C

1X GlycoBuffer 2
50 mM Sodium Phosphate
(pH 7.5 @ 25°C)

Storage Buffer

20 mM Tris-HCl
50 mM NaCl
1 mM EDTA
pH 7.5 @ 25°C

Heat Inactivation

65°C for 10 minutes

Molecular Weight

Theoretical: 35578 kDa

Unit Assay Conditions

Two fold dilutions of IdeZ Protease (IgG-specific) are incubated with 1 μg of human IgG and 1X GlycoBuffer 2 in a 10 µl reaction. The reaction mix is incubated for 15 minutes at 37°C. Separation of reaction products are visualized by SDS-PAGE.

Storage Notes

  • Avoid repeated freeze/thaw cycles.

Product Notes

  1. Store at -20°C for up to two years.
  2. IdeZ Protease is active on human IgG. It can also be used to prepare mouse IgG2a and IgG3 fragments.
    IdeZ Protease will also cleave Fc-fusion proteins, such as Enbrel.
    IdeZ Protease is active on some monkey, rabbit, and sheep IgG.
  3. IdeZ Protease does not cleave mouse IgG1 or IgG2b, rat, porcine, bovineor goat IgG. It also does not cleave non-IgG isotypes including IgA,IgM, IgD and IgE.

FAQs & Troubleshooting

FAQs

  1. What is the advantage of IdeZ over IdeS?
  2. Is IdeZ Protease active in Phosphate Buffered Saline (PBS)?
  3. Can protein A magnetic beads (NEB# S1425S) be used to create Fc and Fab fragment pools after cleavage with IdeZ Protease?
  4. Can IdeZ Protease cleave IgGs other than human?
  5. Can IdeZ Protease cleave Ig isotypes other than IgG?
  6. Can PNGase F be used together with IdeZ Protease under native conditions to deglycosylate the Fc portion of an antibody?
  7. What is the cleavage site for IdeZ Protease (IgG-specific)?

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.

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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