Etanercept Antibody | AbD25940_hIgG4Pro

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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 HRP conjugated Human anti etanercept (clone AbD25940_hIgG4Pro, HCA277P) at a concentration of 2 μg/ml in HISPEC diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate.

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Human anti etanercept specificity titration ELISA

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_hIgG4Pro, 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.

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Etanercept bridging ELISA for pharmacokinetic (PK) assay development

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_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.

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Etanercept bridging ELISA for anti-drug antibody (ADA) assay development

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_hIgG4Pro). Detection was performed using HRP conjugated etanercept. Data are shown as the mean of three measurements.

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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 AbD25940, or clone AbD26183, 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.

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  • Human anti Etanercept:HRP
  • Human anti Etanercept
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-Etanercept Antibody is a recombinant, inhibitory antibody in IgG4 format; it is ideal for bioanalytical assays to measure levels of etanercept and biosimilars. It is recommended for use in a PK bridging ELISA to detect free drug, and may be used as a reference standard in an ADA assay.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD25940_hIgG4Pro
    • Isotype
      IgG4
    2 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA277PEdatasheet pdfdatasheet pdf0.1 mg
      HCA277P
      HCA277Edatasheet pdfdatasheet pdf0.1 mg
      HCA277
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • Human anti etanercept, clone AbD25940_hIgG4Pro 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_hIGg4Pro 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.

        Clone AbD25940_hIgG4Pro 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.
      • Intended Use
      • 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
        Human IgG4 antibody (kappa light chain) selected from the HuCAL® phage display library and expressed in a human cell line - liquid
      • Reconstitution
      • Preparation
        Purified IgG prepared by affinity chromatography on Protein A
        Purified IgG prepared by affinity chromatography on Protein A
      • Preservative Stabilisers
        0.01% Thiomersal
        0.01% Thiomersal
      • Immunogen
        Etanercept
      • Purity
      • 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 0.1mg/ml
        IgG concentration 0.5 mg/ml
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
        Phosphate buffered saline
        Phosphate buffered saline
      • Storage
        Store at -70oC.
        Storage in frost-free freezers is not recommended.
        This product should be stored undiluted
        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.
      • Shelf Life
        12 months from date of despatch
        12 months from date of despatch
      • 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
        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
      • 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
      • 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 NameYesNoMin DilutionMax Dilution
        ELISA
      • Application NameYesNoMin DilutionMax 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
      • Technical Advice
        Recommended protocols and further information about HuCAL recombinant antibody technology can be found in the HuCAL Antibodies Technical Manual
      • Recommended Protocol
      • Recommended Protocol
      • 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
      • ELISA
        This product may be used in a direct or indirect ELISA or as a detection antibody in a bridging ELISA. Protocol: ADA bridging ELISA
      • Immunohistology
      • Immunohistology
      • Histology Positive Control Tissue
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Immunofluorescence
      • Western Blotting
      • Western Blotting
      • Instructions For Use
      • Instructions For Use

      Additional Etanercept Antibody Formats

      Formats Clone Applications Sizes available
      Etanercept Antibody : HRP AbD25940_hIgG4Pro E 0.1 mg
      Etanercept Antibody : Purified AbD25940_hIgG4Pro E 0.1 mg
      • Copyright © 2017 Bio-Rad Antibodies (formerly AbD Serotec)

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Human anti EtanerceptHCA2760.1 mgE
            HCA276
            Human anti EtanerceptHCA2770.1 mgE
            HCA277
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A
            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Human anti Etanercept:HRPHCA277P0.1 mgE
            HCA277P
            Human anti EtanerceptHCA2760.1 mgE
            HCA276
            Human anti EtanerceptHCA2790.1 mgE
            HCA279
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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