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Etanercept Antibody | AbD25939
Human anti etanercept specificity ELISA
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.
Human anti etanercept specificity titration ELISA
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.
Etanercept bridging ELISA for pharmacokinetic (PK) assay development (free drug)
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.
Etanercept bridging ELISA for pharmacokinetic (PK) assay development (total drug)
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.
Human anti etanercept inhibition ELISA
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.
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Product TypeMonoclonal Antibody
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CloneAbD25939
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IsotypeHuCAL Fab monovalent
1 Formats Available
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
Summary
Secondary Antibodies
Negative Isotype Controls
Useful Reagents
Positive Controls
Histology Controls
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References
Reviews
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- 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.
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Intended Use
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Product FormA 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.
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Reconstitution
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PreparationMetal chelate affinity chromatography
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Preservative Stabilisers0.01% Thiomersal
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ImmunogenEtanercept
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Purity
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AffinityThe monovalent intrinsic affinity of AbD25939 was measured as KD = 3.2 nM by real time, label free molecular interaction analysis on immobilized etanercept.
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Approx. Protein ConcentrationsAntibody concentration 0.5 mg/ml
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Reagents In The Kit
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Preparing The Antibody
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Test Principle
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Buffer SolutionPhosphate buffered saline
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StorageStore 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. -
Shelf Life12 months from date of despatch
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AcknowledgementsSold 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. -
Licensed UseFor 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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RegulatoryFor research purposes only
- 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 Yes No 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 AdviceRecommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
- Recommended Protocol
- ELISAThis 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 - Immunohistology
- Histology Positive Control Tissue
- Immunofluorescence
- Western Blotting
- Instructions For Use
Additional Etanercept Antibody Formats
| Formats | Clone | Applications | Sizes available |
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| Etanercept Antibody : Purified | AbD25939 | E | 0.1 mg |
- Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)
Recommended Negative Isotype Control
Useful Reagents
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Human anti Etanercept | HCA279 | 0.1 mg | E | HCA279 | |
| Hispec Assay Diluent | BUF049A | 50 ml | IY | BUF049A |
Recommended Positive Controls
Histology Controls
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Copyright © 2017 Bio-Rad Laboratories, Inc. All rights reserved
Copyright © 2017 Bio-Rad Laboratories, Inc. All rights reserved
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