FabRICATOR® Z Enzyme
FabRICATOR Z Enzyme is supplied as a lyophilized powder with no preservatives added, and is available in 2000 unit vials for digestion of up to 2 mg mouse IgG2a and IgG3.
FabRICATOR Z (IdeZ) is a cysteine protease that digests mouse IgG2a and IgG3 at a specific site below the hinge, generating a homogenous pool of F(ab')2 and Fc fragments after 2 hours incubation.
Detailed Information
FabRICATOR Z digests mouse IgG2a and IgG3 at a specific site below the hinge region, producing a homogenous pool of F(ab')2 and Fc fragments. Most mouse IgG2a are digested by FabRICATOR Z within 2 hours although the enzyme:ab ratio and/or digestion time may need to be optimized for certain antibodies. There is no risk of overdigestion if the incubation time is prolonged. Since FabRICATOR Z digests IgG under physiological reaction conditions, the immunoreactivity is preserved.
FabRICATOR Z is derived from S. equi ssp. zooepidemicus and expressed in E. coli. The enzyme contains a His-tag and the molecular weight is 36 442 Da.
Storage and Stability
The FabRICATOR Z enzyme is supplied lyophilized, and is reconstituted by the addition of water. After reconstitution, FabRICATOR Z is stable for 1 month at +4-8°C. The product is shipped at ambient temperature, and should be stored at -20°C upon arrival.
Unit Definition
One unit digests ≥ 95% of 1µg mouse IgG2a when incubated in 10 mM Tris, 10 mM NaCl, pH 6.5 at 37°C for 2 hours.
FabRICATOR Z, 2000 Units
A0-FRZ-020€450.00Add to cart
How to Order
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Quotations
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Documents
Instructions
Product Specification
Safety Data Sheet
Certificate of Analysis
FAQ and Support
Latest FAQ
An alternative would be to increase the ratio enzyme to antibody (i.e. Follow protocol for mouse IgG). We have not made any specific tests on low concentrations but we have not heard that it would be any problems.
Yes, FabRICATOR can cleave Fc-Fusion proteins. For example Enbrel is effectivelty cleaved by FabRICATOR into Fc and TNFR. Enbrel (etanercept) is a fusion protein composed of the Fc domain of hIgG1 and the p75 tumor necrosis factor receptor (TNFR).
FabRICATOR is very specific and small changes in the sequence (..CPAPELLGGPSVF..) may reduce or completely impair the ability to digest the IgG.
We have tested a few different modified sequences and we can see that the enzyme digest the IgG to some degree to Fab2 and Fc under somewhat modified reaction conditions. We then obtained a mixture of Fc, Fab2 and single cut mAb (one of the HC still undigested).
We used PBS buffer but with greatly reduced salt concentration (5-10 mM instead of 140 mM) which appeared to promote digestion. We also added 10X more enzyme than recommended amount and prolonged incubation time.
It is very difficult to predict the outcome for a specific modification. It might work, but we cannot guarantee it unfortunately.
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