Golimumab antibody | AbD20710
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 golimumab (clone AbD20710) at a concentration of 2 µg/ml, followed by an HRP-labelled anti-His tag in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate.
A microtiter plate was coated over night with golimumab, human TNFα, adalimumab, human IgG1/kappa or human IgG1/lambda at a concentration of 5Umu;g/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti golimumab (clone AbD20710) titrated to the given concentrations in PBST, followed by an HRP-labelled anti-His tag in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements.
A microtiter plate was coated over night with Human anti golimumab (clone AbD20710) at a concentration of 1μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of golimumab. Detection was performed using HRP-conjugated Human anti golimumab (clone AbD20710) at a concentration of 2μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit. Data is presented as the mean of three measurements.
A microtiter plate was coated over night with TNFα at a concentration of 1μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of golimumab (0.3μg/ml) plus increasing concentrations of the inhibitory Type 1 Human anti golimumab (clone AbD20710) or the non-inhibitory Type 2 Human anti golimumab (clone AbD20897, HCA242) was added. Free golimumab, still capable of binding to the TNFα-coated plate, was detected using Mouse anti Human IgG (Fc) CH2 domain:HRP (MCA647P) and QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements.
A microtiter plate was coated over night with Human anti golimumab (clone AbD20710) at a concentration of 1μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of golimumab. Detection was performed using HRP-conjugated Human anti golimumab (clone AbD20692_hIgG1, HCA241) at a concentration of 2μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit. Data is presented as the mean of three measurements.
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E ResetHuman anti Golimumab
- Product Type
- Monoclonal Antibody
- Clone
- AbD20710
- Isotype
- HuCAL Fab monovalent
- Specificity
- Golimumab
Antibody Quality and 
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| Human anti golimumab antibody, clone AbD20710 is a Type 1 anti-idiotypic antibody that recognizes free golimumab but not the drug/tumor necrosis factor alpha (TNFα) complex. Clone AbD20710 inhibits the binding of golimumab to TNFα and can be used to measure free golimumab levels in serum from patients, or for pharmacokinetic (PK) assay development. An affinity matured variant of HCA240 is available with about 2-fold affinity improvement, clone AbD25429 (HCA286). Golimumab, marketed under the brand name Simponi, is a recombinant human IgG1/kappa monoclonal antibody approved for the treatment of rheumatoid arthritis, psoriatic arthritis and ankylosing spondylitis. This therapeutic antibody, directed against TNFα, acts by blocking the binding of TNFα to its receptors, resulting in a down-regulation of the inflammatory response associated with autoimmune diseases. View a summary of all anti-golimumab antibodies |
- Product Form
- A monovalent human recombinant Fab (kappa light chain) selected from the HuCAL phage display library, expressed in E. coli. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain. This antibody is supplied as a liquid.
- Preparation
- Metal chelate affinity chromatography.
- Buffer Solution
- Phosphate buffered saline.
- Preservative Stabilisers
- 0.01% Thiomersal.
- Immunogen
- Golimumab.
- Affinity
- The intrinsic affinity of the monovalent form of Human anti Golimumab antibody, clone AbD20710 is KD = 2.3 nM as measured by real time, label free molecular interaction analysis on immobilized Golimumab.
- 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.
Simponi is a trademark of Janssen Biotech, Inc. in the USA and Merck & Co, Inc. in Europe.
His-tag is a trademark of EMD Biosciences.
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.
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- Synonyms
- Simponi
- 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.
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