Denosumab Antibody | AbD26296

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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 Human Anti-Denosumab Antibody (clone AbD26296) at a concentration of 2 μg/ml and an HRP conjugated anti-DYKDDDDK tag antibody in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.

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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 AbD2629) 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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Human Anti-Denosumab Antibody bridging ELISA for pharmacokinetic (PK) assay development

A microtiter plate was coated overnight with Human Anti-Denosumab Antibody (clone AbD26296) 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 denosumab. Detection was performed using Human Anti-Denosumab:HRP Antibody clone AbD26295_hIgG1 (HCA280P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of four measurements.

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Human Anti-Denosumab Antibody inhibition ELISA

A microtiter plate was coated overnight with recombinant human RANKL at a concentration of 1 μg/ml. After washing and blocking with PBST + 5% BSA, denosumab was added (0.3 μg/ml), spiked with increasing concentrations of each of three Human Anti-Denosumab Antibodies, clone AbD26295_hIgG (HCA280), clone AbD26862_hIgG1 (HCA282), both in monovalent Fab format in this assay, and clone AbD26296 (HCA288). Detection of denosumab was performed using Mouse Anti-Human IgG (Fc) CH2 Domain:HRP Antibody (MCA647P) followed by QuantaBlu Fluorogenic Peroxidase Substrate. Data is shown as the mean of three measurements.

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  • Human anti Denosumab
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    Anti-denosumab antibody is a recombinant, inhibitory anti-idiotypic antibody in monovalent Fab format; it is recommended as capture antibody in a PK bridging ELISA format with HCA280 for detection.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD26296
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA288Edatasheet pdfdatasheet pdf0.1 mg
      HCA288
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • Human Anti-Denosumab Antibody, clone AbD26296 is a paratope specific, anti-idiotypic antibody that specifically recognizes the monoclonal antibody drug 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 the levels of free denosumab and biosimilar products in bioanalytical assays.

        A pair of anti-denosumab antibodies can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody, in monovalent Fab format, is recommended as the capture antibody, paired with an anti-denosumab antibody in full immunoglobulin format, clone AbD26295_hIgG1 (HCA280) as the detection antibody.

        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
        A monovalent human recombinant Fab (kappa 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
        Denosumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of AbD26296 was measured as KD = 7.1 nM by real time, label free molecular interaction analysis on immobilized denosumab
      • 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. 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
        Clone AbD26296 can be used in a direct or indirect ELISA system or as capture antibody for denosumab in a bridging ELISA together with HCA280P (AbD26295_hIgG1) as the detection reagent.
        Protocol: PK bridging ELISA to measure total drug
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Denosumab Antibody Formats

      Formats Clone Applications Sizes available
      Denosumab Antibody : Purified AbD26296 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 Denosumab:HRPHCA280P0.1 mgE
            HCA280P
            Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
            MCA5995P
            LYNX Rapid HRP Antibody Conjugation KitLNK001P1 Conjugation For 400µg AntibodyCJ
            LNK001P
            Human anti DenosumabHCA2800.1 mgE
            HCA280
            Human anti DenosumabHCA2810.1 mgE
            HCA281
            Human anti DenosumabHCA2820.1 mgE
            HCA282
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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