SpyTag antibody | AbD51767kg
A microtiter plate was coated overnight with a Fab fused to either the original version of SpyTag (Fab-Spy-H) or the optimized version 2 (Fab-F-Spy2-H) or 3 of SpyTag (Fab-F-Spy3-H) at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using the anti-SpyTag antibody, clone AbD51767kg (HCA406) or a rabbit anti-DYKDDDDK tag antibody titrated to the given concentration followed by HRP conjugated goat anti-rabbit IgG (H/L) antibody (STAR124P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with SpyCatcher version 2 (H-SpyC2) coupled to a Fab fused to either the original version of SpyTag (Fab-Spy-H) or the optimized version 2 (Fab-F-Spy2-H) or 3 of SpyTag (Fab-F-Spy3-H) at a concentration of 1 μg/ml. An uncoupled Fab fused to SpyTag version 2 (Fab-F-Spy2-H) was used as a control. After washing and blocking with PBST + 5% BSA, detection was performed using the anti-SpyTag antibody, clone AbD51767kg titrated to the given concentration followed by HRP conjugated goat anti-rabbit IgG (H/L) antibody (STAR124P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
10, 15 or 20 ng of a Fab fused to either the original version of SpyTag (Fab-Spy-H) or the optimized version 2 (Fab-F-Spy2-H) or 3 of SpyTag (Fab-F-Spy3-H) were run on a 4-20% Criterion TGX Stain-Free Protein Gel (Bio-Rad #5678095) under denaturing conditions. Proteins were transferred to a 0.2 μm PVDF membrane (Bio-Rad #1704157) and detection was performed using anti-SpyTag antibody, clone AbD51767kg diluted 1:100 in TBST + 1% BSA followed by HRP conjugated goat anti-rabbit IgG (H/L) antibody (STAR124P) and Clarity Western ECL Substrate (Bio-Rad #1705061). M, marker (Bio-Rad #1610374).
10, 15 or 20 ng of E.coli lysate, maltose-binding protein fused to SpyTag version 1 at its N-terminal end (Spy-MBP-His), IgG1 fused to SpyTag version 3 at its C-terminal end (IgG1-Spy3) or SpyCatcher version 3 (SpyCatcher3) were run on a 4-20% Criterion TGX Stain-Free Protein Gel (Bio-Rad #5678095) under denaturing conditions. Proteins were transferred to a 0.2 μm PVDF membrane (Bio-Rad #1704157) and detection was performed using anti-SpyTag antibody, clone AbD51767kg diluted 1:100 in TBST + 1% BSA followed by HRP conjugated goat anti-rabbit IgG (H/L) antibody (STAR124P) and Clarity Western ECL Substrate (Bio-Rad #1705061). M, marker (Bio-Rad #1610374).
anti SpyTag
- Product Type
- Monoclonal Antibody
- Clone
- AbD51767kg
- Isotype
- IgG
- Specificity
- SpyTag
Antibody Quality and Performance Guarantee
| Anti-SpyTag Antibody, clone AbD51767kg is a chimeric antibody comprising a human Fab portion and a rabbit Fc domain, which specifically binds to SpyTag protein with high affinity. SpyTag is a 13 amino acid peptide tag deriving from the second immunoglobulin-like collagen adhesin domain (CnaB2) from the fibronectin binding protein (FbaB) of Streptococcus pyogenes and mainly used for protein ligation in combination with the SpyCatcher (Zakeri et al. 2012).
Anti-SpyTag Antibody binds to the original SpyTag as well as the optimized derivatives SpyTag2 (Keeble et al. 2017) and SpyTag3 (Keeble et al. 2019). It recognizes the SpyTag variants in uncoupled form when fused to both N-terminal or C-terminal end of the target protein. The anti-SpyTag Antibody has been shown to not react with any SpyCatcher variants nor with E.coli lysate. Only residual detection of SpyTag ligated to SpyCatchers was observed in ELISA. |
|
- Target Species
- Synthetic Peptide
- Product Form
- Recombinant chimeric human/rabbit IgG antibody (lambda light chain) selected from the HuCAL phage display library and expressed in a human cell line. In this antibody, the VH-CH1 regions and light chains are human while the Fc region is derived from rabbit IgG. This antibody is supplied as a liquid.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Source
- HKB-11
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.09% Sodium Azide (NaN3)
- Immunogen
- SpyTag version 1 and 2
- Approx. Protein Concentrations
- Antibody concentration 1.0 mg/ml
- 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.
This product is shipped at ambient temperature. It is recommended to aliquot and store at -20°C on receipt. When thawed, aliquot the sample as needed. Keep aliquots at 2-8°C for short term use (up to 4 weeks) and store the remaining aliquots at -20°C.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
Avoid repeated freezing and thawing as this may denature the antibody. Storage in frost-free freezers is not recommended.
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visit the antibody protocols page.
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA |  |
1/10,000 | 1/20,000 |
| Western Blotting | |
1/5,000 | 1/10,000 |
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
- Technical Advice
- For further information about recombinant antibodies incorporating SpyTag technology visit SpyTag and SpyCatcher Products.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Goat anti Rabbit IgG (H/L):HRP | STAR124P | C E WB | 1 mg |
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| Description | Goat anti Rabbit IgG (H/L):HRP | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
| List Price | Your Price | ||||||
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| Description | Hispec Assay Diluent | ||||||
| anti SpyCatcher | HCA379 | E F WB | 0.1 mg |
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| Description | anti SpyCatcher | ||||||
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| SpyTag3 Peptide | BLP086 | IY | 1 mg |
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| Description | SpyTag3 Peptide | ||||||
| HuCAL Fab-F-Spy2-H Negative Control Antibody | TZA003 | E | 0.1 mg |
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| Description | HuCAL Fab-F-Spy2-H Negative Control Antibody | ||||||
HCA406
166414
167714
168759
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