SmartEnzymes FabRICATOR Z workflow

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.

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.
 

  • 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
Mouse IgG2a and IgG3, human IgG1-4, IgG of some classes from monkey, rabbit and sheep
2 h reaction
No need for reducing agents or co-factors
CPAPNLLG / GPSVF (below the hinge)

FabRICATOR® Z Lyophilized

Lyophilized enzyme for below hinge digestion of mouse IgG2a and IgG3

FabRICATOR® Z Immobilized

Immobilized enzyme for below hinge digestion of mouse IgG2a and IgG3 in spin columns

FabRICATOR® Z Fab2 Kit

Immobilized enzyme and affinity resin for below hinge digestion of mouse IgG2a and IgG3 and purification of fragments

About FabRICATOR® Z

FabRICATOR Z workflow

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.

FabRICATOR® Z Lyophilized

Lyophilized enzyme for below hinge digestion of mouse IgG2a and IgG3.

FabRICATOR Z Lyophilized is available as a lyophilized powder in 2000 unit vials for digestion of 2 mg mouse IgG2a and IgG3.
 

  • Flexible format for method development
  • Possible to combine enzymes for improved efficiency
  • Amenable to automated workflows
  • Ready to use – just add water!

Available Products

FabRICATOR Z Lyophilized 2000 unitsLyophilized enzyme for below hinge digestion of 2 mg mouse IgG2a and IgG3A0-FRZ-020607.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One unit digests ≥ 95% of 1µg mouse IgG2a when incubated in 10 mM sodium phosphate, 10 mM NaCl, pH 6.5 at 37°C for 2 hours.

Content and Storage

FabRICATOR Z Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, the enzyme 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.

Product Specifications

Safety Data Sheet

Certificate of Analysis

FabRICATOR® Z Immobilized (formerly known as FragIT Z)

Immobilized enzyme for below hinge digestion of mouse IgG2a and IgG3 in spin columns.

The FabRICATOR Z Immobilized spin columns contain the FabRICATOR Z enzyme covalently coupled to agarose beads, for digestion of mouse IgG2a and IgG3 without contaminating the final preparation with enzyme.

The Microspin columns are available for digestion of 2 x 0.5 mg, 5 x 0.5 mg, or 10 x 0.5 mg mouse IgG2a or IgG3.
 

  • Easy-to-use spin columns
  • Increased enzyme-to-substrate ratio for improved digestion efficiency
  • No enzyme in the final sample

Available Products

FabRICATOR Z Immobilized Microspin 2 x 0.5 mgImmobilized enzyme for below hinge digestion of 2 x 0.5 mg mouse IgG2a and IgG3 in spin columnsA0-FZ6-010456.00Buy / Request a Quote
FabRICATOR Z Immobilized Microspin 5 x 0.5 mgImmobilized enzyme for below hinge digestion of 5 x 0.5 mg mouse IgG2a and IgG3 in spin columnsA0-FZ6-0251,028.00Buy / Request a Quote
FabRICATOR Z Immobilized Microspin 10 x 0.5 mgImmobilized enzyme for below hinge digestion of 10 x 0.5 mg mouse IgG2a and IgG3 in spin columnsA0-FZ6-0501,695.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One spin column contains sufficient FabRICATOR Z-coupled agarose beads to digest 0.5 mg mouse IgG2a or IgG3.

Content and Storage

FabRICATOR Z Immobilized is supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the product!

Product Specifications

Safety Data Sheet

Certificate of Analysis

FabRICATOR® Z Fab2 Kit (formerly known as FragIT Z Kit)

Immobilized enzyme and affinity resin for below hinge digestion of mouse IgG2a and IgG3 and purification of fragments.

FabRICATOR Z Fab2 Kit consists of spin columns of FabRICATOR Z Immobilized for antibody digestion, and spin columns of CaptureSelect™* Fc resin for purification of fragments. One FabRICATOR Z spin column contains sufficient FabRICATOR Z-coupled agarose beads to process 0.5 mg mouse IgG2a and IgG3.
 

  • Purification of homogenous F(ab’)2 fragments from mouse IgG2a and IgG3
  • Fast - pure fragments in under two hours
  • Reliable workflow for preparative applications

Available Products

FabRICATOR Z Fab2 Kit Microspin 0.5 mgImmobilized enzyme and affinity resin for below hinge digestion of 0.5 mg mouse IgG2a and IgG3 and purification of fragmentsA2-FZ2-005514.00Buy / Request a Quote
FabRICATOR Z Fab2 Kit Microspin 5 x 0.5 mgImmobilized enzyme and affinity resin for below hinge digestion of 5 x 0.5 mg mouse IgG2a and IgG3 and purification of fragmentsA2-FZ2-0251,367.00Buy / Request a Quote

For information on how to order, visit Place an Order or contact us directly at order@genovis.com.

Unit Definition

One spin column contains sufficient FabRICATOR Z-coupled agarose beads to digest 0.5 mg mouse IgG2a or IgG3.

Content and Storage

The FabRICATOR Z Immobilized spin columns and CaptureSelect™* spin columns are supplied in 20% ethanol, shipped cold, and should be stored at +4-8°C upon arrival. Do not freeze the products!

Instructions

Product Specifications

Safety Data Sheet

Certificate of Analysis

* Thermo Scientific™ CaptureSelect™ resin from Thermo Fisher Scientific Inc. and its subsidiaries. Thermo Fisher and CaptureSelect are trademarks of Thermo Fisher Scientific Inc. and its subsidiaries.

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 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.

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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