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Cat. No.: IZP-1k (for 1KU)

Cat. No.: IZP-5k (for 5KU)

Cat. No.: IZP-20k (for 20KU)

Description

IdeZ Protease (IgG-specific) is an Immunoglobulin-degrading Enzyme derived from Streptococcus equi subspecies zooepidemicus. IdeZ Protease exhibits extremely high substrate specificity, recognizing only specific species and types of IgG, and cleaving at specific sites below the antibody hinge region, resulting in the hydrolysis of IgG into intact F(ab')2 fragments and Fc fragments. As a tool enzyme, IdeZ Protease is primarily used for structural characterization analysis of antibody-based drugs or antibody fusion proteins. It can be utilized to prepare antibodies or antibody fragments containing only F(ab')2 fragments, as well as to remove interference from heavy chains during Western detection following immunoprecipitation for the accurate detection of target proteins, among other applications.

IdeZ Protease effectively cleaves human IgG1, IgG2, IgG3, and IgG4, as well as IgG from rabbit, rat, monkey, sheep, humanized and chimeric IgG, and Fc fusion proteins such as Enbrel. However, IdeZ Protease cannot cleave mouse IgG1/IgG2b, pig, cow, or goat IgG, nor non-IgG isotypes of immunoglobulins, including IgA, IgM, IgD, and IgE. Compared to IdeS Protease, IdeZ Protease exhibits significantly improved enzymatic activity for cleaving rat and mouse IgG2a and IgG3.

Protein Information

• Molecular Weight: Approximately 33 kDa.
• Applications: IdeZ Protease is used for structural characterization analysis of antibody-based drugs or antibody fusion proteins. It is employed for enzymatic cleavage to generate antibodies or antibody fragments containing only F(ab')2 fragments or Fc fragments. It can also be used to remove interference from heavy chains during Western detection following immunoprecipitation of target proteins.
• Source: Purified from recombinant E. coli strain carrying the IdeZ Protease gene.
• Enzymatic Activity: IdeZ Protease (IgG-specific) is a recombinant antibody-specific protease that recognizes all human, rabbit, rat, sheep, and monkey IgG subclasses. It cleaves specifically at a single recognition site below the hinge region, yielding a homogenous pool of F(ab')2 and Fc fragments. IdeZ Protease (IgG-specific) exhibits greater efficiency in cleaving murine IgG2a compared to IdeS Protease.
• Unit Definition: One unit is defined as the amount of enzyme required to cleave >95% of 1µg of human IgG in 15 minutes at 37°C in a total reaction volume of 10µl.
• Purity: The purity of IdeZ Protease is greater than 95% as determined by SDS-PAGE. It does not contain any other proteases apart from IdeZ Protease, nor does it contain DNA endonucleases or exonucleases, RNAse, or phosphatases.
• Enzyme Storage Buffer: 20mM Tris-HCl, 50mM NaCl, 1mM EDTA (pH 7.5 @25°C).
• IdeZ Protease Dilution Buffer: 20mM Tris-HCl, 50mM NaCl, 1mM EDTA (pH 7.5 @25°C).
• 10X Reaction Buffer: 500mM Sodium Phosphate (pH 7.5 @25°C).
• Inactivation or Inhibition: IdeZ Protease can be inactivated by incubation at 65°C for 10 minutes.

Precautions

• IdeZ Protease should be stored in an icebox or on an ice bath during use and promptly placed at -20°C after each use.
• IdeZ Protease exhibits variations in enzymatic efficiency when cleaving IgG from different species. When cleaving specific species' IgG, it is recommended to explore the appropriate reaction incubation time and enzyme dosage. IdeZ Protease has lower cleavage efficiency for mouse IgG2a and IgG3, so it is advised to increase the amount of IdeZ Protease (recommended starting point: 5 to 10 times the usual dosage) or extend the reaction incubation time (2 hours to overnight) to achieve sufficient cleavage of mouse IgG2a and IgG3.
• IdeZ Protease exhibits the highest activity in neutral or near-neutral pH buffer systems, and the salt concentration in the cleavage system should not be excessively high. It is recommended to use the provided 10X Reaction Buffer for the cleavage reaction. When using buffers outside the pH range of 6-8 (e.g., acetate buffer), the incubation time or enzyme dosage may need to be optimized based on the specific conditions.
• This product is intended for scientific research purposes by professionals only and is not to be used for clinical diagnosis or treatment, food or drug purposes, or stored in regular residential settings.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling this product.

Storage

The minimum shelf life is 2 years at -20°C. Avoid repeated freeze-thaw cycles. It is recommended to aliquot the product appropriately upon first use and store it in the freezer for future use.

Only for research and not intended for treatment of humans or animals

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