Etanercept Antibody | AbD25939

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Etanercept Antibody | AbD25939 gallery image 1

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.

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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 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.

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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.

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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.

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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 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.

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  • Human anti Etanercept
(Rated 0.0 out of 5 based on 0 customer reviews)
    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
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA276Edatasheet pdfdatasheet pdf0.1 mg
      HCA276
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      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.
      • Intended Use
      • 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.
      • Reconstitution
      • Preparation
        Metal chelate affinity chromatography
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Etanercept
      • Purity
      • 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
      • Reagents In The Kit
      • Preparing The Antibody
      • Test Principle
      • Buffer Solution
        Phosphate buffered saline
      • 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.
      • Shelf Life
        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.
      • 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

      • 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
      • Recommended Protocol
      • 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
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Etanercept Antibody Formats

      Formats Clone Applications Sizes available
      Etanercept Antibody : Purified AbD25939 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 EtanerceptHCA2790.1 mgE
            HCA279
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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