Etanercept antibody | AbD25939
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 etanercept (clone AbD25939) at a concentration of 2 μg/ml followed by a HRP conjugated anti-DYKDDDDK tag antibody 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 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human anti etanercept (clone AbD25939) titrated to the given concentrations in PBST, with a HRP conjugated anti-DYKDDDDK tag antibody in HISPEC 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) 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_hIgG4Pro, 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 Human anti etanercept (clone AbD25939), 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 AbD26183_hIgG1, HCA279P) 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 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 AbD25939, or clone AbD25940, (HCA277) 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.
Human anti Etanercept
Anti-Etanercept Antibody is a recombinant, non-inhibitory monovalent Fab antibody; it is ideal for bioanalytical assays to measure levels of etanercept and biosimilars. It is recommended as capture antibody in a PK bridging ELISA with HCA277P or HCA279P.
- Product Type
- Monoclonal Antibody
- Clone
- AbD25939
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA276 | E | 0.1 mg |
|
Human anti etanercept, clone AbD25939 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 AbD25939 does not inhibit the binding of etanercept to its target tumor necrosis factor alpha (TNFα). The antibody binds to an epitope outside the TNFα binding site and can be used to measure the levels of etanercept and biosimilar products (e.g. Etacept, Intacept) in bioanalytical assays.
Our anti-etanercept antibodies can be used to develop pharmacokinetic (PK) bridging assays to measure either free drug or total drug. This antibody, in monovalent Fab format, is recommended as the capture antibody. For an assay to detect free drug, use with the inhibitory antibody, clone AbD25940_hIgG4Pro (HCA277P) for detection; for an assay to detect total drug use with p75-Fc-fusion specific clone AbD26183_hIgG1 (HCA279P) for detection.
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.
Our anti-etanercept antibodies can be used to develop pharmacokinetic (PK) bridging assays to measure either free drug or total drug. This antibody, in monovalent Fab format, is recommended as the capture antibody. For an assay to detect free drug, use with the inhibitory antibody, clone AbD25940_hIgG4Pro (HCA277P) for detection; for an assay to detect total drug use with p75-Fc-fusion specific clone AbD26183_hIgG1 (HCA279P) for detection.
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 Details
- Product Form
- A monovalent human recombinant Fab (lambda 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
- Etanercept
- Affinity
- The monovalent intrinsic affinity of AbD25939 was measured as KD = 3.2 nM by real time, label free molecular interaction analysis on immobilized etanercept.
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- Store at +4oC or at -20oC if preferred.
Storage in frost-free freezers is not recommended.
This product should be stored undiluted. Avoid repeated freezing and thawing as this may denature the antibody. Should this product contain a precipitate we recommend microcentrifugation before use. - Guarantee
- 12 months from date of despatch
More Information
- Acknowledgements
- Sold under license of U.S. Patents 6753136, 7785859 and 8273688 and corresponding patents. This antibody was developed by Bio-Rad, Zeppelinstr. 4, 82178 Puchheim, Germany.
Enbrel® is a registered trademark of Immunex Corporation.
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.
- Regulatory
- For research purposes only
Applications of Etanercept antibody
| 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 or indirect ELISA or as capture antibody for etanercept.
Protocol: PK bridging ELISA to measure free drug
Protocol: PK bridging ELISA to measure total drug
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Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Etanercept | HCA277 | E | 0.1 mg |
|
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| Human anti Etanercept:HRP | HCA277P | E | 0.1 mg |
|
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| Human anti Etanercept | HCA279 | E | 0.1 mg |
|
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| Human anti Etanercept:HRP | HCA279P | E | 0.1 mg |
|
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