Etanercept antibody | AbD25940_hIgG4_Pro
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 HRP conjugated Human anti etanercept (clone AbD25940_hIgG4_Pro, HCA277P) at a concentration of 2 μg/ml in HISPEC diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate.
A microtiter plate was coated overnight with etanercept, TNFR2, Fc control or infliximab at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HRP conjugated Human anti etanercept (clone AbD25940_hIgG4_Pro, HCA277P) titrated to the given concentrations in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data for binding to etanercept and TNFR2 are shown as the mean of three measurements.
A microtiter plate was coated overnight with Human anti etanercept (clone AbD25939, HCA276) at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of etanercept. Detection was performed using HRP conjugated Human anti etanercept (clone AbD25940_hIgG4_Pro, HCA277P) at a concentration of 2 μg/ml 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 etanercept 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 etanercept (clone AbD25940_hIgG4_Pro). Detection was performed using HRP conjugated etanercept. Data are shown as the mean of three measurements.
A microtiter plate was coated overnight with human TNFα at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of etanercept (0.3 μg/ml) plus increasing concentrations of Human anti etanercept (clone AbD25940_hIgG4_Pro, or clone AbD26183_hIgG1, HCA279) in the monovalent Fab format was added. Free etanercept, still capable of binding to the human TNFα coated plate, was detected using Mouse anti Human IgG (Fc) CH2 Domain:HRP (MCA647P) and QuantaBlu™ fluorogenic peroxidase substrate. Data are shown as the mean of three measurements.
Filter by Application:
E ResetHuman anti Etanercept:HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD25940_hIgG4_Pro
- Isotype
- IgG4
- Specificity
- Etanercept
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| Human anti etanercept, clone AbD25940_hIgG4_Pro is an antibody that specifically recognizes etanercept as well as the extracellular ligand-binding portion of the 75 kDa (p75) human tumor necrosis factor receptor 2 (TNFR2). Clone AbD25940_hIgG4_Pro inhibits the binding of etanercept to its target tumour necrosis factor alpha (TNFα). The antibody binds to free etanercept but not to etanercept bound to its target TNFα. The antibody binds to an epitope which is also part of the TNFα binding site and can be used to measure the levels of etanercept and biosimilar products (e.g. Etacept, Intacept) in bioanalytical assays. CloneAbD25940_hIgG4_Pro is a fully human recombinant monoclonal antibody with an IgG4 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay. Additionally the antibody can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in full immunoglobulin format, is recommended as the detection antibody, paired with the antibody clone AbD25939 (HCA276) in monovalent Fab format, as the capture antibody. Etanercept (Enbrel®) is a dimeric fusion protein consisting of the extracellular ligand-binding portion of the human 75 kDa (p75) TNFR2 linked to the Fc portion of human IgG1. The Fc component of etanercept contains the CH2 domain, the CH3 domain and hinge region, but not the CH1 domain of IgG1. It is used to treat autoimmune diseases by interfering with TNF, acting as a TNF inhibitor. It binds to both TNFα and TNFβ (Lymphotoxin A). View a summary of all anti-etanercept antibodies. |
- Product Form
- Human IgG4 antibody (kappa light chain) selected from the HuCAL? phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) - liquid
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Etanercept
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=2.4 nM by real time, label-free molecular interaction analysis on immobilized etanercept
- Approx. Protein Concentrations
- IgG concentration 0.1 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.
Enbrel is a registered trademark of Immunex Corporation.
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 and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- ELISA
- This product may be used in a direct ELISA or as a detection antibody in a bridging ELISA.
Protocol: PK bridging ELISA to measure free drug
| 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 Etanercept | HCA276 | E | 0.1 mg |
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| Description | Human anti Etanercept | ||||||
| Human anti Etanercept | HCA277 | E | 0.1 mg |
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| Description | Human anti Etanercept | ||||||
- Synonyms
- ENBREL®
- 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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