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FabRICATOR®Z IdeZ Antibody Cleavage

FabRICATOR® Z (IdeZ)

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.

 

FabRICATOR Z is available in three different product formats:

Product Information

FabRICATOR Z is an enzyme for preparation of F(ab’)2 from mouse subclasses IgG2a and IgG3. FabRICATOR Z digests mouse IgG2a and IgG3 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. For digestion of human IgGs, FabRICATOR® is recommended.

FabRICATOR Z digests IgG at a specific site and there is no risk of overdigestion if the incubation time is prolonged. FabRICATOR Z digests IgG at mild conditions, thus preserving the immunoreactivity. 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.

Key Characteristics

  • Mouse IgG2a and IgG3, IgG from monkey, rabbit and sheep
  • 2 h reaction
  • No need for reducing agents or co-factors
  • CPAPNLLG / GPSVF (below the hinge)

Applications

  • Digest mouse IgG2a and IgG3

Product Formats

FabRICATOR® Z Enzyme

The FabRICATOR Z enzyme supplied as a lyophilized powder. The enzyme is available in 2000 unit vials for digestion of up to 2 mg mouse IgG2a or IgG3.

FragIT™ Z (Immobilized FabRICATOR Z)

FragIT Z is FabRICATOR Z (IdeZ) enzyme immobilized on agarose beads, allowing for mouse IgG2a and IgG3 digestion without contaminating the final preparation with FabRICATOR Z enzyme.

FragIT™ Z kit

FragIT Z Kit easily generates and purifies F(ab')2 and Fc/2 fragments from mouse IgG2a and IgG3. The kit consists of two parts; FragIT Z spin columns and affinity spin columns for purification of F(ab'2) fragments.
 


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FabRICATOR-Z Applications

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FAQ and Support

Popular FAQ

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 FabUSLOUS 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.

It is possible to elute the Fc fragments from the CaptureSelect column using a low pH buffer, for example a 0.1M Glycine buffer pH=3.0. The pH must immediately after elution be adjusted to neutral pH by adding 0.1x elution volume of 1M Tris pH=8.0.

 

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