Obinutuzumab antibody | AbD50319ia
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 AbD50319ia (HCA403) directly conjugated to HRP at a concentration of 2 μg/mL in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P).
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 AbD50319ia (HCA403) directly conjugated to HRP, titrated to the given concentrations in HISPEC diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data are shown as the mean of three measurements. HRP conjugation was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P).
Figure A. PE conjugated obinutuzumab pre-incubated for 1h with Human Anti-Obinutuzumab Antibody, clone AbD50319ad (HCA403, red) or recombinant Human IgG1 Kappa Allotype G1m3 Antibody, 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 AbD50319ad (HCA403, red) or recombinant Human IgG1 Kappa Allotype G1m3 Antibody, 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 obinutuzumab at a concentration of 1 µg/ml. After washing and blocking with PBST + 5% BSA, PBST with 10% human serum was added spiked with increasing concentrations of Human Anti-Obinutuzumab Antibody, clone AbD50319ia (HCA403, black), clone AbD50514ia (HCA404, red) or clone AbD50104ia (HCA405, blue). Detection was performed using HRP conjugated obinutuzumab at a concentration of 2 µg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit (LNK002P). Data are shown as the mean of three measurements.
Human anti Obinutuzumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD50319ia
- Isotype
- IgG1
- Specificity
- Obinutuzumab
Antibody Quality and Performance Guarantee
HCA403
| Human Anti-Obinutuzumab Antibody, clone AbD50319ia 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 AbD50319ia (HCA403) is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a surrogate positive control or calibrator in an anti-drug antibody (ADA) assay. This antibody has a monovalent intrinsic affinity of 0.562nM and is one of the three antibodies offered for ADA assays. Antibody HCA404 has an affinity of 1.318nM and HCA405 has an affinity of 15.607nM. 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
- Human IgG1 antibody (lambda light chain) selected from Bio-Rad’s human antibody phage display libraries and expressed in a human cell line. 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
- Obinutuzumab
- Affinity
- The monovalent intrinsic affinity of Human anti-Obinutuzumab Antibody, clone AbD50319ia was measured as KD = 0.562 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.
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 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 and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual.
- ELISA
- Clone AbD50319ia may be used to develop and calibrate immune response assays to measure the anti-drug antibody (ADA) response in patient sera together with clone AbD50514ia (HCA404) and clone AbD50104ia (HCA405).
Protocol: ADA 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 | 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 | ||||||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | Log in | |||
| List Price | Your Price | ||||||
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| Description | LYNX Rapid HRP Antibody Conjugation Kit | ||||||
| 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 | ||||||
| Human anti Obinutuzumab:HRP | TZA049P | E | 0.1 mg | Log in | |||
| List Price | Your Price | ||||||
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| Description | Human anti Obinutuzumab:HRP | ||||||
- Synonyms
- Gazyvaro
HCA403
167082
If you cannot find the batch/lot you are looking for please contact our technical support team for assistance.
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