FragIT™ Z kit
FragIT Z kit is used for the preparation and purification of F(ab')2 and Fc fragments from mouse IgG2a and IgG3. The kit consists of spin columns of FragIT Z (Immobilized FabRICATOR Z) for antibody digestion and spin columns of CaptureSelect™* Fc resin for affinity purification of the Fc fragment.
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.
FragIT Z is a resin with the FabRICATOR Z (IdeZ) enzyme covalently coupled to agarose beads for subunit fragmentation of mouse IgG2a and IgG3 without contaminating the final preparation with enzyme. The FragIT Z kit consists of FragIT Z resin and CaptureSelect™ columns containing multispecies Fc affinity matrix. The used ligand is directed towards the Fc domain of IgG enabling purification of IgG from several species, including human, mouse, bovine, rabbit, rat, goat, horse and sheep.
The FragIT Z kit is available for digestion of 0.5 mg or 5 x 0.5 mg mouse IgG2a or IgG3.
Storage and Stability
The FragIT Z 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!
Buffers
FragIT Z can be used with all commonly used buffers with pH ranging from 6.0 to 8.0. Optimization may be required. Note that the FabRICATOR Z enzyme is irreversibly inactivated below pH 5.0.
Unit Definition
One spin column contains sufficient FabRICATOR Z-coupled agarose beads to digest 0.5 mg mouse IgG2a or IgG3.
*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.
How to Order
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Quotations
Please do not hesitate to contact us if you need a quotation or have inquiries on price of bulk quantities.
Documents
Instructions
Product Specifications
Safety Data Sheet
FAQ and Support
Latest FAQ
FabRICATOR® cleave the IgG molecule in one specific site in the lower hinge region of the anitbody leaving a F(ab')2 fragment and a Fc-fragment.
Human IgG1: HTCPPCPAPELLG⬇GPSVF
Human IgG2: VECPPCPAPP_VA⬇GPSVF
Human IgG3: PPCPRCPAPELLG⬇GPSVF
Human IgG4: PHAHHAQAPEFLG⬇GPSVF
You can indeed elute the Fc fragments from the CaptureSelect column using a low pH buffer commonly used for example during protein A/G purification of antibodies. We have successfully used 0.1 M glycine buffer pH 2.6-2.8 to release the Fc fragments. Be sure to adjust pH back to normal using a 1 M Tris-HCl buffer or similar directly after isolation of the Fc fragments from the CaptureSelect column.
To use protein A or MabSelect Sure (GE) for removal of Fc as a platform after digestion with FabRICATOR or FragIT is problematic. Therefore we have chosen CaptureSelect in the FragIT Kit for removal of Fc fragments. CaptureSelect is much more specific for Fc. Both protein A and MabSelect Sure (GE) may have affinity for Fab and the degree of affinity is antibody dependent. We have many customer reports on this subject and they usually comes back to us for the CaptureSelect columns for the above reason.
Find all FAQ here
Patent and Disclaimer
FabRICATOR®
Genovis AB hold a license for exclusive worldwide use of biotechnical industrial applications of IdeS covered by any patents derived from International publication WO03051914, including granted US Patent No US7666582. Other aspects of FabRICATOR®technology are encompassed by pending patent applications in the name of Genovis AB.
The trademark FabRICATOR® is the property of Genovis AB.
For research use only. Not intended for any animal or human therapeutic diagnostic use.