Denosumab Antibody | AbD26296_hIgG1

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Human Anti-Denosumab Antibody 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-Denosumab Antibody (clone AbD26296_hIgG1, HCA281) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate. HRP conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit®.

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Human Anti-Denosumab Antibody specificity titration ELISA

A microtiter plate was coated overnight with denosumab, human IgG2/kappa or human IgG1/lambda at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Denosumab Antibody (clone AbD26296, here as his-tagged, monovalent Fab fragment) titrated to the given concentrations in PBST, followed by detection with Mouse Anti-Penta Histidine Tag:HRP Antibody (MCA5995P) and QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.

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

A microtiter plate was coated overnight with denosumab at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, 10% human serum was added spiked with increasing concentrations of Human Anti-Denosumab Antibody, clone AbD26295_hIgG1 (HCA280, black line; KD = 1.8 nM), clone AbD26296_hIgG1 (HCA281, red line; KD = 7.1 nM) or clone AbD26862_hIgG1 (HCA282, blue line; KD = 0.8 nM). Detection was performed using HRP conjugated denosumab followed by QuantaBlu Fluorogenic Peroxidase. HRP conjugation of the denosumab used in this assay was performed using a LYNX Rapid HRP Conjugation Kit®. Data is shown as the mean of three measurements.

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  • Human anti Denosumab
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-denosumab antibody is a recombinant, inhibitory, anti-idiotypic antibody in IgG1 format for use in bioanalysis and immunogenicity testing of denosumab and biosimilars. It can be used as detection antibody in a PK bridging ELISA and as a reference standard in an ADA bridging ELISA.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD26296_hIgG1
    • Isotype
      IgG1
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA281Edatasheet pdfdatasheet pdf0.1 mg
      HCA281
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Human Anti-Denosumab Antibody, clone AbD26296_hIgG1 is a paratope specific anti-idiotypic antibody that specifically recognizes the free human monoclonal antibody denosumab. The antibody does not recognize free RANKL (receptor activator of nuclear factor kappa-B ligand) or denosumab in complex with human RANKL and can be used to measure free denosumab and biosimilar products in bioanalytical assays.

        Clone AbD26296_IgG1 is a fully human recombinant monoclonal antibody with IgG1 isotype and is suitable as a reference standard in an anti-drug antibody (ADA) assay.

        Denosumab (Prolia, Xgeva) is a fully human monoclonal antibody (IgG2/kappa) for the treatment of osteoporosis, treatment-induced bone loss, bone metastases, multiple myeloma, and giant cell tumor of bone. The drug specifically binds to human RANKL a protein that acts as the primary signal to promote bone removal/resorption and prevents RANKL from activating its receptor, RANK, on the surface of osteoclasts and their precursors.

        View a summary of all anti-denosumab antibodies.
      • Intended Use
      • Product Form
        Human IgG1 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
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Denosumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=7.1 nM by real time, label-free molecular interaction analysis on immobilized denosumab
      • Approx. Protein Concentrations
        IgG 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. Prolia and Xgeva are registered trademarks of Amgen Inc.
      • 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 antibody is fully human and can be used as a reference standard in an ADA assay.
        Protocol: ADA bridging ELISA
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Denosumab Antibody Formats

      Formats Clone Applications Sizes available
      Denosumab Antibody : Purified AbD26296_hIgG1 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 DenosumabHCA2800.1 mgE
            HCA280
            Human anti DenosumabHCA2820.1 mgE
            HCA282
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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