Trastuzumab Antibody | AbD25279

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Human anti trastuzumab (drug/target complex) specificity titration ELISA

A microtiter plate was coated over night with human IgG1/kappa, human ErbB2, alemtuzumab or trastuzumab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, the human ErbB2/trastuzumab complex was formed by adding 2 μg/ml trastuzumab to the wells coated with human ErbB2, detection was performed using Human anti trastuzumab (clone AbD25279) titrated to the given concentrations in PBST+10% human serum, followed by HRP labeled mouse anti His tag (MCA5995P) in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data is presented as the mean of three measurements

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Human anti trastuzumab (drug/target complex) 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 trastuzumab (clone AbD25279) at a concentration of 2 μg/ml, followed by HRP labeled mouse anti DYKDDDDK (MCA4764P) in HISPEC assay diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate

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Trastuzumab antigen capture ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with human ErbB2 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 trastuzumab. Detection was performed using human anti trastuzumab (clone AbD25279) at a concentration of 2 μg/ml, followed by HRP labeled mouse anti-His tag antibody (MCA5995P) in HISPEC diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data is presented as the mean of three measurements

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  • Human anti Trastuzumab (Drug/Target Complex)
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-trastuzumab drug-target complex antibody specifically recognizes the trastuzumab- HER2/neu receptor complex exclusively and detects trastuzumab or biosimilars only when bound to the HER2/neu receptor. This antibody in monovalent Fab format is suitable for a PK antigen capture format assay.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD25279
    • Isotype
      HuCAL Fab bivalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA263Edatasheet pdfdatasheet pdf0.1 mg
      HCA263
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • Human anti trastuzumab drug/target complex, clone AbD25279, is a bivalent Fab antibody that specifically recognizes trastuzumab when in a complex with its target, the extracellular domain of the human epidermal growth factor receptor 2 (HER2, also known as ErbB2). It does not recognize the free drug or unbound HER2. The antibody can be used in pharmacokinetic (PK) antigen capture format assays to measure trastuzumab that has been captured via immobilized HER2. It is ideal for bioanalytical assays to measure the level of drug or biosimilar in complex with the HER2 target.

        Trastuzumab (trade name Herceptin®) is a drug used in the treatment of HER2 positive breast cancer and other HER2 overexpressing cancers including HER2-positive metastatic cancers of the gastrointestinal tract. Trastuzumab binds to the HER2 (or c-erbB2) proto-oncogene, an EGF receptor-like protein found on 20-30% of breast cancer cells. The binding leads to antibody mediated (complement mediated) killing of the HER2 positive cells.

        View a summary of all anti-trastuzumab antibodies
      • Intended Use
      • Product Form
        A bivalent human recombinant Fab selected from the HuCAL® phage display library, expressed in E. coli. This Fab fragment is bivalent by dimerization of the bacterial alkaline phosphatase fusion protein. The antibody is tagged with a DYKDDDDK tag and a HIS-tag (HHHHHH) at the C-terminus of the antibody heavy chain.
      • Reconstitution
      • Preparation
        Metal chelate affinity chromatography
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Trastuzumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=31 nM by real time, label-free molecular interaction analysis on immobilized trastuzumab/ErbB2 complex.
      • 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 6,300,064, 6,696,248, 6,708,484, 6,753,136, European Patent 0,859,841 and corresponding patents. This antibody was developed by Bio-Rad, a wholly owned subsidiary of Bio-Rad Laboratories Inc., Zeppelinstr. 4, 82178 Puchheim, Germany.
        Herceptin® is a registered trademark of Genentech, 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 product may be used as a detection reagent in a direct or an indirect ELISA for trastuzumab captured by immobilized human ErbB2.
        Protocol: PK antigen capture ELISA to measure bound drug exclusively
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Trastuzumab Antibody Formats

      Formats Clone Applications Sizes available
      Trastuzumab Antibody : Purified AbD25279 E 0.1 mg
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            Rat anti DYKDDDDK Tag:HRPMCA4764P0.1 mgE, P, WB
            MCA4764P
            Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
            MCA5995P
            LYNX Rapid HRP Antibody Conjugation KitLNK002P3 Conjugations For 400µg AntibodyCJ
            LNK002P
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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