FabRICATOR® Z – Below Hinge Digestion of Mouse IgG

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 a two-hour incubation.

SmartEnzymes™

The enzyme is mostly used for preparative generation of mouse IgG2a and IgG3 subunits, but it is also active on IgG from human, monkey, rabbit and sheep.

Why FabRICATOR® Z?

  • Specific – one digestion site below the hinge
  • Efficient for digestion of mouse IgG2a and IgG3
  • Generates a homogenous pool of fragments within two hours
  • Available immobilized in ready-to-use spin columns

  • Target

    Mouse IgG2a and IgG3, human IgG1-4, IgG of some classes from monkey, rabbit and sheep

  • Time

    2 h reaction

  • Box

    No need for reducing agents or co-factors

  • Digestion Site

    CPAPNLLG / GPSVF (below the hinge)

Product Formats

About FabRICATOR® Z

FabRICATOR Z is a cysteine protease for digestion of mouse IgG2a and IgG3. It digests the IgG at a specific site below the hinge region, generating intact F(ab’)2 and Fc fragments.

Most mouse IgG2a and IgG3 are digested by FabRICATOR Z within 2 hours, although the enzyme:antibody ratio and/or digestion time may need to be optimized for certain antibodies. FabRICATOR Z is also active on IgG from monkey, rabbit and sheep. For digestion of human IgG, we recommend using FabRICATOR.

Since FabRICATOR Z digests IgG at a specific site, there is no risk of overdigestion if the incubation time is prolonged. FabRICATOR Z digests IgG in sodium phosphate buffer with pH ranging from 6.0-8.0, but is most active at pH 6.5 and at low NaCl concentrations.

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.

A method to detect the FabRICATOR Z enzyme is available. Learn more about Anti- FabRICATOR Z, a goat polyclonal antibody.

Citations

FAQ and Support

FabRICATOR MagIC is suitable for using in automated systems that can handle magnetic beads. On the applications page you can read about how it’s used in a KingFisher™ workflow.

Unfortunately, FabRICATOR Z does not digest rat IgG2a. If generating Fab fragments is an option, we suggest using FabULOUS enzyme that digests rat IgG2a. FabULOUS does not have a single specific cleavage site, but a primary digestion site, so there can be more than one digestion site. The digestion site(s) for rat IgG2a using FabULOUS is not determined, but the generated fragments are Fab and Fc.

We can only guarantee optimal digestion for one-time use. We do not have a cleaning or regeneration protocol to provide. However, depending on the antibody and the following application the column can be reused. We do not recommend using with different antibodies due to the risk for contamination of carry-over from previous sample. If column reuse is desired store the column in 10-20% ethanol at +4-8°C.

No, FabRICATOR Z only digests mouse IgG2a (….CPAPNLLG / GPSVF.. ) and IgG3 (…CPAPNILG / GPSVF…) at one specific site below the hinge. If Fab fragments are an option, we recommend our FabULOUS enzyme that digest mouse IgG1 above the hinge. FabULOUS requires light reducing conditions for optimal activity. If strong reducing conditions are used, this may likely cause reduction of the interchain disulfides.

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