FabULOUS™
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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 Lyophilized is available as a lyophilized powder in 2000 unit vials for digestion of 2 mg IgG.
Why FabULOUS™ Lyophilized?
- Flexible format for method development
- Possible to combine enzymes for improved efficiency
- Amenable to automated workflows
- Ready to use – just add water!
Available Products
FabULOUS™ Lyophilized 2000 units
Lyophilized enzyme for above hinge digestion of 2 mg IgG
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 human IgG1 when incubated in PBS with 5 mM DTT or TCEP, pH 7.4 at 37°C for 1 hour.
Buffers
FabULOUS digests IgG in commonly used buffers with pH ranging from 6.5 to 8.0 and best activity is obtained at 37°C. The enzyme requires reducing agent in the digestion reaction and depending on the reducing conditions, there is a chance that the interchain thiols will be reduced.
Content and Storage
FabULOUS Lyophilized is reconstituted by addition of water. No preservatives added. After reconstitution, FabULOUS is stable for at least 1 month at +4-8°C. The product is shipped cold and should be stored at -20°C upon arrival.
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
Will the reducing conditions required for digestion impact the interchain disulfide bond integrity?
Depending on what reducing agent and what concentration you use you can obtain intact Fab and not reduce the interchain disulfide bond. The light chain (LC) and heavy chain (HC) will not dissociate under native conditions due to hydrogen/hydrophobic forces. Before you analyze your samples with SDS-PAGE you must remove the cysteine otherwise the Fab will be reduced to Fd and LC during sample preparation for the SDS-PAGE. This can be done by a simple buffer exchange.
How can I stop the FabULOUS reaction?
FabULOUS enzyme reaction is inhibited with iodoacetamide, iodo acetic acid or lowering pH below 3.
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…).
Is FabULOUS working on human IgG4?
Yes, but a longer incubation time is needed.
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.
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