FabULOUS™
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FabULOUS™ Fab Kit
Lyophilized enzyme and affinity resin for hinge-region digestion of murine IgG and purification of fragments
FabULOUS (SpeB) is a cysteine protease that digests in the hinge region of IgG from several different species and subclasses, generating Fab and Fc fragments.
FabULOUS Fab Kit contains one vial of FabULOUS enzyme with 2000 units for digestion of 2 mg murine IgG, and 4 CaptureSelect™ LC-Kappa (mur)* microspin columns for affinity purification of fragments.
The affinity matrix recognizes the LC-Kappa domain of murine IgG antibodies, which enables purification of Fab fragments. Each microspin column contains sufficient material to purify 0.5 mg IgG.
Why FabULOUS™ Fab Kit?
- Purification of Fab fragments from murine IgG
- Fast – pure fragments in under two hours
- Reliable workflow for preparative applications
Available Products
FabULOUS™ Fab Kit 2 mg
Lyophilized enzyme and affinity resin for above hinge digestion of 2 mg murine IgG and purification of fragments
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Product Information
Unit Defintion
One unit digests ≥ 95% of 1 µg of human IgG1 in PBS pH 7.4 with 5 mM DTT or TCEP at 37˚C for 1 hour. This is also valid for 1 µg of mouse IgG1 in PBS with 30-50 mM L-cysteine as reducing agent.
Content and Storage
FabULOUS Fab Kit is shipped cold.The FabULOUS Lyophilized vials should be stored at -20°C upon arrival.
The microspin columns containing CaptureSelect™* LC-Kappa (mur) resin should be stored at +4-8°C. Do not freeze the spin columns!
After reconstitution, the FabULOUS enzyme is stable for 1 month at +4-8°C.
Resources
Instruction for Use
Product Specifications
Safety Data Sheet
Certificate of Analysis
Find your certificate of analysis »Performance Activity
Preparation of pure, homogeneous mouse IgG Fab fragments with no residual enzyme by combining FabULOUS digestion and affinity purification.
Citations
Powered by BiozFAQ and Support
Is FabULOUS working on human IgG4?
Yes, but a longer incubation time is needed.
Can I digest Fc-fusion proteins using FabULOUS (SpeB)?
Yes, FabULOUS should be able to digest a Fc fusion protein as long as you have not fused the protein too close to the cleavage site. FabULOUS digests human IgG1 just above the hinge (…KTHT / CPPCP…).
I work with mouse IgG2a containing LC-lambda, can I use another affinity resin for purification of the Fab?
Yes. There is the possibility to exchange the CaptureSelect LC-kappa (mur) columns to columns containing CaptureSelect LC-lambda (mouse). This resin is specific for mouse IgG. Please contact support@genovis.com if this is of interest.
Does FabULOUS digest all human IgG subclasses above the hinge?
FabULOUS is a cysteine protease that digests IgG in the hinge region. A primary digestion site for human IgG1 is above the hinge (…KTHT / CPPCP…). The enzyme is also active on human IgG2 and IgG3, digestion site not determined but Fab and Fc is generated. FabULOUS digests human IgG4 primarily at a site below the hinge using significantly longer incubation, generating Fab’ due to the reducing conditions. The possibility of other, secondary digestions sites is antibody dependent
Does FabULOUS need reducing condition to work?
Yes, FabULOUS requires reduced condition for activity and reaction is performed in the presence of 1-5 mM DTT or TCEP at neutral pH.
Legal & Disclaimers
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Genovis products are intended for research use only. They are not intended to be used for therapeutic or diagnostic purposes in humans or animals. All goods and services are sold subject to Genovis’ General Terms and Conditions of Sale.
* 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.
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