BiSpyCatcher2
BiSpyCatcher2:Biotin
- Product Type
- Recombinant Protein
- Specificity
- BiSpyCatcher2
BiSpyCatcher2 (H-BiSpyC2) is a 29 kDa homodimer of SpyCatcher2 (TZC001) bridged with a flexible linker. Each SpyCatcher2 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. BiSpyCatcher2 contains a His6-tag at the N-terminus. To avoid deamidation of asparagine, a N105D mutation was introduced (Hentrich et al. 2020) for each of the two Catchers. 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 BiSpyCatcher2 undiluted. All the different formats of BiSpyCatcher2 are compatible with Bio-Rad's recombinant HuCAL® antibodies with a SpyTag2 at the C-terminus of the heavy chain. Download BiSpyCatcher coupling protocol BiSpyCatcher2 (TZC002) is available in two modified versions, BiSpyCatcher2-CYS (TZC002CYS) and BiSpyCatcher-CYS3 (TZC002CYS3), which contain one or three cysteine residues, respectively, for site-specific conjugation by the user. BiSpyCatcher2 (TZC002) is also available conjugated to biotin (TZC002B) and HRP (TZC002P). BiSpyCatcher2-CYS can dimerize by formation of a disulfide bond via the free cysteines. Before conjugation to these cysteines, the BiSpyCatcher2-CYS must be reduced e.g., by addition of 5mM DTT and incubation for 1 hr at room temperature, followed by a fast DTT removal step, e.g., by size exclusion chromatography, as DTT can interfere with the conjugation chemistry. BiSpyCatcher2-CYS3 contains DTT to avoid oligomerization through disulfide bond formation. Immediately before conjugation, to avoid oxidation and disulfide bond formation, carry out a fast DTT removal step, e.g., by size exclusion chromatography. View all available Catchers |
- Target Species
- Protein/peptide tag
- Product Form
- Purified recombinant protein conjugated to Biotin - liquid
- Preparation
- Recombinant protein expressed in E. coli and purified by affinity chromatography
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.0095% MIT
- Purity
- ≥90% determined by SDS-PAGE under reducing conditions and visualized by coomassie blue staining
- Approx. Protein Concentrations
- 80 µM (2.4 mg/ml)
- Protein Molecular Weight
- Predicted molecular weight of the purified Catcher is 29469 Da
The molecular weight specified excludes conjugation and will vary upon conjugation depending on the molecular weight of the label - Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- 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.
His-tag is a registered trademark of EMD Biosciences - 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.
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 |
---|---|---|---|
Immunoassay |
- Instructions For Use
-
TZC002B
View BiSpyCatcher Coupling Protocol
Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
---|---|---|---|---|---|---|---|
SpyTag3 Peptide | BLP086 | IY | 1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | SpyTag3 Peptide | ||||||
anti SpyCatcher | HCA379 | E F WB | 0.1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | anti SpyCatcher | ||||||
BiSpyCatcher2 | TZC002 | IY | 1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | BiSpyCatcher2 | ||||||
BiSpyCatcher2-CYS | TZC002CYS | IY | 1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | BiSpyCatcher2-CYS | ||||||
BiSpyCatcher2-CYS3 | TZC002CYS3 | IY | 1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | BiSpyCatcher2-CYS3 | ||||||
BiSpyCatcher2:HRP | TZC002P | IY | 1 mg | Log in | |||
List Price | Your Price | ||||||
Log in | |||||||
Description | BiSpyCatcher2:HRP |
Source Reference
-
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.
References for BiSpyCatcher2
-
Keeble, A.H. et al. (2017) Evolving Accelerated Amidation by SpyTag/SpyCatcher to Analyze Membrane Dynamics.
Angew Chem Int Ed Engl. 56 (52): 16521-16525. -
Hentrich, C. et al. (2021) Periplasmic expression of SpyTagged antibody fragments enables rapid modular antibody assembly.
Cell Chem Biol. 28 (6): 813-824.e6.
TZC002B
162614 163734 165890 168275If you cannot find the batch/lot you are looking for please contact our technical support team for assistance.
Please Note: All Products are "FOR RESEARCH PURPOSES ONLY"
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