Powered by BiozAbove Hinge Digestion of IgG
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.
The prepared Fab fragments can for example be used in affinity studies, studies of Fab glycosylation and structural studies.
- Active on IgG from multiple species and subclasses – digests in the upper-hinge region
- Fast - generates Fab and Fc fragments within one hour
- Efficient for digestion of mouse IgG1
FabULOUS™ Lyophilized
Lyophilized enzyme for above hinge digestion of IgGFabULOUS™ Fab Kit
Lyophilized enzyme and affinity resin for above hinge digestion of murine IgG and purification of fragmentsAbout FabULOUS™
FabULOUS (SpeB) is an enzyme for digestion of IgG in the hinge region. Human, mouse, rat, goat and sheep IgG are digested by FabULOUS, yielding Fab and Fc fragments. The primary digestion site on human IgG1 is …KTHT / CPPCPAPE…
FabULOUS is derived from Streptococcus pyogenes and is expressed in E. coli. The molecular weight of FabULOUS is 29 kDa and the enzyme contains a His-tag.
FabULOUS™ Citations
FabULOUS™ Lyophilized
Lyophilized enzyme for above hinge digestion of IgG.
FabULOUS Lyophilized is available as a lyophilized powder in 2000 unit vials for digestion of 2 mg IgG.
- Flexible format for method development
- Possible to combine enzymes for improved efficiency
- Amenable to automated workflows
- Ready to use – just add water!
Available Products
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Generate Intact Fab and Fc Fragments
Unit Definition
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.
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.
Buffers and Reducing Agents
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.
Instructions
Product Specifications
Safety Data Sheet
Certificate of Analysis
FabULOUS™ Fab Kit
Lyophilized enzyme and affinity resin for above hinge digestion of murine IgG and purification of 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.
- Purification of Fab fragments from murine IgG
- Fast - pure fragments in under two hours
- Reliable workflow for preparative applications
Available Products
For information on how to order, visit Place an Order or contact us directly at order@genovis.com.
Generate Intact Fab from Mouse IgGUnit Definition
One unit digests ≥ 95% of 1µg human or mouse IgG when incubated in 10 mM Tris, 150 mM NaCl pH 7.4 at 37°C for 1 hour.
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.
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
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.
No, FabULOUS Fab kit contains CaptureSelect LC-kappa (mur) spin columns with selective affinity for murine LC-kappa. If you need to generate pure Fab from human IgG1 you can use either FabALACTICA Fab Kit or GingisKHAN Fab Kit. If you have a different subclass of human IgG, please contact support@genovis.com for guidance.
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
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…).
Yes, FabULOUS requires reduced condition for activity and reaction is performed in the presence of 1-5 mM DTT or TCEP at neutral pH.
Support