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|Human IgG4-Pro-FcSpyCatcher3 (hIgG4-Pro-FcSpyC3) contains a SpyCatcher3 (Keeble et al. 2019) fused to the hinge region, CH2 and CH3 of human IgG4-Pro (Hentrich et al. 2020), resulting in a 76.5 kDa protein for the disulfide-bridged dimer. The IgG4-Pro sequence has a mutation S228P in the core hinge region that prevents the formation of IgG4 half molecules. Each SpyCatcher3 subunit can form a stable covalent isopeptide bond with a second protein that includes a SpyTag. The reaction occurs spontaneously on mixing, is rapid, irreversible, high-yielding and shows good specificity.
The reaction is robust at pH 5 to 8, at temperatures from +4°C to +37°C, in various buffer conditions (Ca2+/Mg2+ not needed) and in the presence of detergents. The reaction also occurs inside cells (in vivo). Since the speed of the coupling reaction is concentration dependent, it is recommended to use hIgG4-Pro-FcCatcher3 undiluted.
Human IgG4-Pro-FcSpyCatcher3 is compatible with Bio-Rad's recombinant HuCAL antibodies with a SpyTag2 at the C-terminus of the heavy chain.
Download FcSpyCatcher coupling protocol
View all available Catchers
- Target Species
- Protein/peptide tag
- Product Form
- Purified recombinant protein - Liquid
- Recombinant protein expressed in mammalian cell line and purified by affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.1% ProClin 950
- ≥90% determined by SDS-PAGE under reducing conditions and visualized by coomassie blue staining
- Approx. Protein Concentrations
- 80 µM (6.1 mg/ml)
- Protein Molecular Weight
- Predicted 76514 Da
- For research purposes only
- 12 months from date of despatch
- ProClin is a trademark of The Dow Chemical Company (“Dow”) or an affiliated company of
This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the protein. Should this product contain a precipitate we recommend microcentrifugation before use.
|Application Name||Verified||Min Dilution||Max Dilution|
- Instructions For Use
- View FcSpyCatcher Coupling Protocol
Antibody Characterization Reference
Hentrich, C. et al. (2021) Periplasmic expression of SpyTagged antibody fragments enables rapid modular antibody assembly.
Cell Chem Biol. 28 (6): 813-824.e6. Cell Chem Biol. 28 (6): 813-824.e6.
Zakeri, B. et al. (2012) Peptide tag forming a rapid covalent bond to a protein, through engineering a bacterial adhesin.
Proc Natl Acad Sci U S A. 109 (12): E690-7. Proc Natl Acad Sci U S A. 109 (12): E690-7.
Keeble, A.H. et al. (2019) Approaching infinite affinity through engineering of peptide-protein interaction.
Proc Natl Acad Sci U S A. 116(52):26523-33. Proc Natl Acad Sci U S A. 116(52):26523-33.
- Licensed Use
- For in vitro research purposes. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.
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