Obinutuzumab antibody | AbD51296
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-Obinutuzumab Antibody, clone AbD51296ad (TZA049) at a concentration of 2 μg/ml followed by an HRP-labelled anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated overnight with obinutuzumab, rituximab, infliximab, human IgG1 kappa control, and human IgG1 lambda control at a concentration of 5 µg/ml. After washing and blocking with PBST + 5% BSA, detection was performed using Human Anti-Obinutuzumab Antibody, clone AbD51296ad (TZA049) titrated to the given concentrations in PBST followed by an HRP-labelled anti-DYKDDDDK tag antibody in HISPEC diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Figure A. PE conjugated obinutuzumab pre-incubated for 1h with Human Anti-Obinutuzumab Antibody, clone AbD51296ad (TZA049, red) or recombinant Human IgG1 Kappa Allotype G1m3, clone AbD18705_hIgG1 isotype control (HCA192G, grey) in the monovalent Fab format in ratio 1:1. Figure B. PE conjugated obinutuzumab pre-incubated for 1h with Human Anti-Obinutuzumab Antibody, clone AbD51296ad (TZA049, red) or recombinant Human IgG1 Kappa Allotype G1m3, clone AbD18705_hIgG1 isotype control (HCA192G, grey) in the monovalent Fab format in ratio 5:1. All experiments were performed on a Daudi cell line expressing CD20 and incubated for 1 hour with the specific antibody or antibody mix.
A microtiter plate was coated overnight with Human Anti-Obinutuzumab Antibody, clone AbD51167ad (TZA048) 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 obinutuzumab. Detection was performed using HRP conjugated Human Anti-Obinutuzumab Antibody, clone AbD51296pap (TZA049P) at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements.
Human anti Obinutuzumab:HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD51296
- Isotype
- Fab antibody
- Specificity
- Obinutuzumab
Antibody Quality and Performance Guarantee
TZA049P
| Human Anti-Obinutuzumab Antibody, clone AbD51296 is a paratope specific, inhibitory, anti-idiotypic antibody (Type 1) that specifically recognizes the free human monoclonal antibody drug obinutuzumab, and its biosimilar products. It does not recognize the drug target, the B cell surface marker CD20, nor obinutuzumab in complex with CD20. This antibody can be used to measure free obinutuzumab levels in serum from patients. Clone AbD51296 is available as a monovalent Fab antibody incorporating a SpyTag at the C-terminus end of the antibody heavy chain for conversion by the end user into alternative formats using any of the SpyCatchers available in our catalog under product code TZA049. It is also available as a bivalent Fab antibody already conjugated to HRP via coupling to the HRP conjugated BiSpyCatcher2 (TZC002P) for use in direct ELISA under product code TZA049P. A pair of anti-obinutuzumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug; clone AbD51296 is recommended as a detection antibody, paired with Human Anti-Obinutuzumab Antibody, clone AbD51167ad (TZA048) as the capture reagent. Obinutuzumab (trade name Gazyvaro) is a humanized monoclonal antibody of the IgG1/kappa isotype that specifically targets the extracellular loop of CD20 when expressed at the surface of non malignant and malignant pre-B and mature B-cells. It does not bind CD20 expressed on hematopoietic stem cells, pro-B cells, normal plasma cells or other normal tissue. In non-clinical studies, obinutuzumab induces direct cell death and mediates antibody dependent cellular cytotoxicity (ADCC) and antibody dependent cellular phagocytosis (ADCP) through recruitment of FcɣRIII positive immune effector cells. In vivo, obinutuzumab mediates a low degree of complement dependent cytotoxicity (CDC). Obinutuzumab is indicated for the combination treatment of patients with chronic lymphocytic leukemia and follicular lymphoma. View a summary of all anti-obinutuzumab antibodies. |
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- Product Form
- A bivalent human recombinant Fab (kappa light chain) generated by coupling of the monovalent SpyTagged Fab TZA049 with a HRP conjugated BiSpyCatcher version 2 (TZC002P). The coupled antibody has two DYKDDDDK tags and three His6-tags. It contains SpyTag3 peptide (BLP086) in 5-fold molar excess to block any unreacted Catcher sites. 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
- Obinutuzumab
- Affinity
- The monovalent intrinsic affinity of Human anti-Obinutuzumab Antibody, clone AbD51296pap was measured as KD = 0.293 nM by real time, label-free molecular interaction analysis on immobilized obinutuzumab.
- 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.
Gazyvaro is a registered trademark of Genentech/ Roche and Biogen. - 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 frozen.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
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 AbD51296pap may be used as the detection antibody together with AbD51167ad (TZA048) as the capture antibody for obinutuzumab in a PK bridging ELISA.
Protocol: PK bridging ELISA
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
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| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Obinutuzumab | HCA403 | E | 0.1 mg |
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| Description | Human anti Obinutuzumab | ||||||
| Human anti Obinutuzumab | HCA404 | E | 0.1 mg |
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| Description | Human anti Obinutuzumab | ||||||
| Human anti Obinutuzumab | HCA405 | E | 0.1 mg |
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| Description | Human anti Obinutuzumab | ||||||
| Human anti Obinutuzumab | TZA048 | E | 0.1 mg | Log in | |||
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| Description | Human anti Obinutuzumab | ||||||
| Human anti Obinutuzumab | TZA049 | E | 0.1 mg | Log in | |||
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| Description | Human anti Obinutuzumab | ||||||
- Synonyms
- Gazyvaro
TZA049P
167087
If you cannot find the batch/lot you are looking for please contact our technical support team for assistance.
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