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Cat. No.: IZPP-1k (for 1KU)

Cat. No.: IZPP-5k (for 5KU)

Cat. No.: IZPP-20k (for 20KU)

Description

IdeZ Protease (IgG-specific, Powder) 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 33kDa.
• Applications: IdeZ Protease is used for structural characterization of antibody-based drugs or antibody fusion proteins. It cleaves antibodies specifically at a single recognition site below the hinge region, generating homogeneous pools of F(ab')2 fragments and Fc fragments. It is also used to remove heavy chains that interfere with target protein detection in immunoprecipitation followed by Western blotting.
• Source: Purified from recombinant E.coli strain carrying the IdeZ Protease gene.
• Enzyme 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 homogeneous pool of F(ab')2 and Fc fragments. IdeZ Protease (IgG-specific) exhibits higher cleavage efficiency for murine IgG2a compared to IdeS Protease.
• Activity Definition: One unit is defined as the amount of enzyme required to cleave >95% of 1µg of human IgG in a total reaction volume of 10µl, at 37°C for 15 minutes.
• Purity: The purity is >95% as determined by SDS-PAGE. It does not contain any other proteases, DNA endonucleases or exonucleases, RNAse, or phosphatases.
• Enzyme Storage Buffer: Lyophilized from 50mM Sodium Phosphate, 150mM NaCl (pH6.6 @25°C).
• IdeZ Protease Dilution Buffer: 50mM Sodium Phosphate, 150mM NaCl (pH6.6 @25°C).
• 10X Reaction Buffer: 500mM Sodium Phosphate (pH7.5 @25°C).
• Inactivation or Inhibition: IdeZ Protease can be inactivated by incubating at 65°C for 10 minutes.

Precautions

• When using IdeZ Protease, it should be stored in a cooler or on ice, and after use, it should be immediately placed at -20°C for storage.
• IdeZ Protease exhibits significant variations in cleavage efficiency among different species' IgG. When cleaving specific species' IgG, it is recommended to adjust the reaction incubation time and enzyme amount accordingly. IdeZ Protease has lower cleavage efficiency for mouse IgG2a and IgG3. It is advised to increase the amount of IdeZ Protease (starting with a 5 to 10-fold higher amount) or extend the reaction incubation time (2 hours to overnight) to achieve sufficient cleavage of mouse IgG2a and IgG3.
• IdeZ Protease exhibits maximum activity in neutral pH 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 10X Reaction Buffer provided with this product for enzyme cleavage reactions. When using buffers outside the pH range of 6-8 (such as acetate buffer), the incubation time or enzyme amount should be optimized according to the specific circumstances.
• This product is intended for scientific research purposes by professionals only. It should not be used for clinical diagnosis or treatment, food or drug purposes, and should not be stored in regular residential areas.
• For your safety and health, please wear laboratory attire and disposable gloves when handling.

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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