Tocilizumab Antibody | AbD21345_hIgG1

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Human anti tocilizumab specificity ELISA

A microtiter plate was coated over night 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 tocilizumab (clone AbD21345_hIgG1) 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 tocilizumab specificity titration ELISA

A microtiter plate was coated over night with human IgG1/kappa, human IgG1/lambda or tocilizumab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using HRP-conjugated Human anti tocilizumab (clone AbD21345_hIgG1) titrated to the given concentrations in PBST, followed by QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements

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Tocilizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti tocilizumab (clone AbD21362, HCA252) 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 tocilizumab. Detection was performed using HRP-conjugated Human anti tocilizumab (clone AbD21345_hIgG1) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements. 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 tocilizumab bridging ELISA for Anti-Drug Antibody (ADA) assay development

A microtitre plate was coated over night with tocilizumab 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 different Human anti tocilizumab clones, AbD21338_hIgG1 (HCA253), AbD21364_hIgG1(HCA254), AbD21346_hIgG1 (HCA255) and AbD21345_hIgG1 (HCA256). Detection was performed using HRP-conjugated tocilizumab. Data is presented as the mean of two measurements. HRP-conjugation of tocilizumab used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit®

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Human anti tocilizumab inhibits the binding of tocilizumab to human interleukin 6 receptor (IL-6R)

A microtiter plate was coated over night with human interleukin 6 receptor at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of tocilizumab (0.3 μg/ml) plus increasing concentrations of the monovalent Fab fragment of the inhibitory Human anti tocilizumab clone AbD21345_hIgG (HCA256) or the non-inhibitory clone AbD22155_hIgG1 (HCA257) was added. Free tocilizumab, still capable of binding to the human interleukin 6 receptor-coated plate, was detected using Mouse anti Human IgG (Fc) CH2 Domain:HRP (MCA647P) and QuantaBlu fluorgenic peroxidase substrate. Data is presented as the mean of three measurements

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  • Human anti Tocilizumab
(Rated 0.0 out of 5 based on 0 customer reviews)
    Anti-tocilizumab antibody is a recombinant, neutralizing anti-idiotypic antibody in fully human IgG1 format, suitable for bioanalytical assays for tocilizumab and biosimilars. It is recommended as a reference standard with lower affinity for an ADA bridging ELISA.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD21345_hIgG1
    • Isotype
      IgG1
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA256Edatasheet pdfdatasheet pdf0.1 mg
      HCA256
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
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      • Human anti tocilizumab, clone AbD21345_hIgG1 is a Type 1 anti-idiotypic antibody that specifically recognizes free tocilizumab but not the drug/Interleukin-6 Receptor (IL-6R) complex. Clone AbD21345_hIgG1 inhibits the binding of tocilizumab to IL-6R and can be used to measure free tocilizumab levels in serum from patients. This clone can also be used for pharmacokinetic (PK) assay development or in the development and calibration of anti-drug antibody (ADA) assays.

        Tocilizumab, marketed under the brand name RoActemra® in Europe and Actemra® in North America, is a humanized IgG1/kappa monoclonal antibody approved for the treatment of moderate to severe rheumatoid arthritis and for the treatment of systemic juvenile idiopathic arthritis. This therapeutic antibody, directed against IL-6R, acts by blocking the binding of IL-6R (both the soluble form, sIL-6R, and the membrane bound form, mIL-6R) to IL-6, thus impeding the pro-inflammatory properties of this cytokine.

        Clone AbD21354_hIgG1 is one of a range of Type 1 (HCA252), (HCA253), (HCA254), (HCA255) and Type 2 (HCA257) anti-idiotypic antibodies to tocilizumab available from Bio-Rad and forms part of a wider and expanding portfolio of anti-idiotypic antibodies that have been developed for the detection of biotherapeutics.
      • Intended Use
      • Product Form
        Human IgG1 antibody selected from the HuCAL® phage display library and expressed in a human cell line. This antibody is supplied as a liquid.
      • Reconstitution
      • Preparation
        Purified IgG prepared by affinity chromatography on Protein A
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Tocilizumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD = 15 nM by real time, label free molecular interaction analysis on immobilized tocilizumab.
      • 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.

        Actemra and RoActemra are registered trademarks of Hoffman-La Roche and Chugai Pharmaceuticals co., Ltd.
      • 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 ELISA or as a detection reagent in a bridging ELISA together with HCA252 as the capture reagent.
        Protocol: PK bridging ELISA to measure free drug
        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 Tocilizumab Antibody Formats

      Formats Clone Applications Sizes available
      Tocilizumab Antibody : Purified AbD21345_hIgG1 E 0.1 mg
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        Recommended Negative Isotype Control

          Useful Reagents

            DescriptionProduct CodePack SizeApplicationsList PriceQuantity
            LYNX Rapid HRP Antibody Conjugation KitLNK005P1 Conjugation For 20mg AntibodyCJ
            LNK005P
            LYNX Rapid HRP Antibody Conjugation KitLNK001P1 Conjugation For 400µg AntibodyCJ
            LNK001P
            LYNX Rapid HRP Antibody Conjugation KitLNK003P1 Conjugation For 4mg AntibodyCJ
            LNK003P
            LYNX Rapid HRP Antibody Conjugation KitLNK006P3 Conjugations For 40µg AntibodyCJ
            LNK006P
            LYNX Rapid HRP Antibody Conjugation KitLNK002P3 Conjugations For 400µg AntibodyCJ
            LNK002P
            LYNX Rapid HRP Antibody Conjugation KitLNK004P5 Conjugations For 4mg AntibodyCJ
            LNK004P
            Human anti TocilizumabHCA2520.1 mgE *
            HCA252
            Human anti TocilizumabHCA2530.1 mgE
            HCA253
            Human anti TocilizumabHCA2540.1 mgE
            HCA254
            Human anti TocilizumabHCA2550.1 mgE
            HCA255
            Human anti TocilizumabHCA2570.1 mgE
            HCA257
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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