MMAE antibody | AbD50348
A microtiter plate was coated overnight with various antigens at a concentration of 5 μg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using Human Anti-MMAE Antibody, clone AbD50348ad (TZA057) at a concentration of 2 μg/ml in PBST followed by an HRP conjugated Anti-DYKDDDDK tag Antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with brentuximab vedotin, MMAE coupled to Human Transferrin (MMAE-TRF), MMAF coupled to Human Transferrin (MMAF-TRF), MMAD coupled to Human Transferrin (MMAD-TRF) and brentuximab (MCA6149) at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using Human Anti-MMAE Antibody, clone AbD50348ad (TZA057) titrated to the given concentrations in PBST followed by an HRP conjugated anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with brentuximab vedotin at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, a pre-incubated mixture of 0.5 µg/ml Human Anti-MMAE Antibody, clone AbD50348ad (TZA057) in the monovalent Fab format plus increasing concentrations of free MMAE toxin(black), free MMAF toxin (red) or free MMAD toxin (blue) was added. Free Human Anti-MMAE Antibody, clone AbD50348ad (TZA057), still capable of binding to the brentuximab vedotin coated plate, was detected using an HRP conjugated Anti-DYKDDDDK tag Antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human Anti-MMAE Antibody, clone AbD50347ad (TZA056) at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, 10% human serum was added spiked with increasing concentrations of brentuximab vedotin. Detection was performed using HRP conjugated Human Anti-MMAE Antibody, clone AbD50348pap (TZA057P) at a concentration of 0.5 µg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human anti MMAE
- Product Type
- Monoclonal Antibody
- Clone
- AbD50348
- Isotype
- Fab antibody
- Specificity
- MMAE
Antibody Quality and Performance Guarantee
| Human Anti-MMAE Antibody, clone AbD50348 recognizes the Monomethyl Auristatin E (MMAE) toxin, an antimitotic agent which inhibits cell division by blocking the polymerization of tubulin.
Because of its toxicity, MMAE is not used as a drug itself, but is rather linked to monoclonal antibodies to form an antibody-drug conjugate through a cathepsin-cleavable linker that is stable in the extracellular fluid. Upon binding into the target cell, the linker is cleaved by cathepsin, realeasing the toxin in the cell and activating the antimitotic mechanism. This antibody is tagged with a SpyTag2 at the C-terminus of the Fab antibody heavy chain, enabling the user to couple it to a SpyCatcher reagent for conversion to alternative formats in less than one hour. View a summary of SpyCatcher products Human anti-MMAE Antibody, clone AbD50348 has been shown to recognize the MMAE toxin in both free and antibody-conjugated forms. It has also been shown to cross-react with the MMAE closely related toxin MMAF and MMAD. It does not recognize unbound antibody drugs. Clone AbD50348 can therefore be used to measure the level of several MMAE bound drug in serum from patients. View a summary of all MMAE antibodies |
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- Product Form
- A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL® phage display library, expressed in a proprietary E. coli strain. The antibody is tagged with a DYKDDDDK tag, a SpyTag version 2 (VPTIVMVDAYKRYK) and a His6-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied as a liquid.
- Preparation
- Metal chelate affinity chromatography
- Source
- E.coli
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.0095% MIT
- Immunogen
- Monomethyl Auristatin E (MMAE)
- Affinity
- The monovalent intrinsic affinity of AbD50348ad was measured as KD = 5 nM by real time, label free molecular interaction analysis on immobilized brentuximab vedotin.
- Approx. Protein Concentrations
- Antibody concentration 0.5 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.
His-tag is a registered trademark of EMD Biosciences. - Licensed Use
- For in vitro research purposes and for commercial applications for the provision of in vitro testing services to support preclinical and clinical drug development. Any re-sale in any form or any other commercial application needs a written agreement with Bio-Rad.
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 |  |
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
- Recommended protocols for coupling a Fab antibody with a SpyTag to a SpyCatcher can be found at SpyTag and SpyCatcher Products, and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- Clone AbD50348ad can be used as the detection antibody for MMAE toxin in its free and also in its coupled format eg. brentuximab vedotin, in a PK ELISA together with TZA056 as the capture antibody.
Protocol: PK bridging ELISA
| 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 | ||||||
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Brentuximab Vedotin | HCA349 | E | 0.1 mg |
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| Description | Human anti Brentuximab Vedotin | ||||||
| Human anti Brentuximab Vedotin | HCA350 | E | 0.1 mg |
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| Description | Human anti Brentuximab Vedotin | ||||||
| Human anti Brentuximab Vedotin | HCA351 | E | 0.1 mg |
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| Description | Human anti Brentuximab Vedotin | ||||||
| Human anti Brentuximab Vedotin | HCA352 | E | 0.1 mg |
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| Description | Human anti Brentuximab Vedotin | ||||||
| Human anti CD30 (Brentuximab Biosimilar) | MCA6149 | E | 0.2 mg |
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| Description | Human anti CD30 (Brentuximab Biosimilar) | ||||||
| Human anti MMAE | TZA056 | E | 0.1 mg |
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| Description | Human anti MMAE | ||||||
| Human anti MMAE:HRP | TZA057P | E | 0.1 mg |
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| Description | Human anti MMAE:HRP | ||||||
TZA057
167540
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