Tocilizumab Antibody | AbD21362

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Tocilizumab Antibody | AbD21362 gallery image 1

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 Human anti tocilizumab (clone AbD21362) at a concentration of 2 μg/ml followed by an HRP conjugated Mouse anti Histidine tag antibody in HISPEC diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate

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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 Human anti tocilizumab (clone AbD21362) titrated to the given concentrations in PBST, followed by followed by an HRP conjugated Mouse anti Histidine tag antibody in HISPEC diluent (BUF049A) and QuantaBlu™ fluorogenic peroxidase substrate. Data presented is 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) 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 AbD21338_hIgG1, HCA253) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) followed by QuantaBlu™ fluorogenic peroxidase substrate. Data presented is 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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Tocilizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti tocilizumab (clone AbD21362) 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 AbD21364_hIgG1, HCA254) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) followed by QuantaBlu™ fluorogenic peroxidase substrate. Data presented is 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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Tocilizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti tocilizumab (clone AbD21362) 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 AbD21346_hIgG1, HCA255) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) followed by QuantaBlu™ fluorogenic peroxidase substrate. Data presented is 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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Tocilizumab Antibody | AbD21362 gallery image 6

Tocilizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti tocilizumab (clone AbD21362) 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, HCA256) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) followed by QuantaBlu™ fluorogenic peroxidase substrate. Data presented is the mean of three measurements. HRP-conjugation of the antibody used in this assay was performed using a LYNX Rapid HRP Antibody Conjugation Kit ®

Enlarge
Tocilizumab Antibody | AbD21362 gallery image 7

Tocilizumab bridging ELISA for Pharmacokinetic (PK) assay development

A microtiter plate was coated over night with Human anti tocilizumab (clone AbD21362) 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 AbD22155_hIgG1, HCA257) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) followed by QuantaBlu fluorogenic peroxidase substrate. Data presented is 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 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 inhibitory Human anti tocilizumab clone AbD21362 (HCA252) or the non-inhibitory monovalent Fab fragment of 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™™ fluorogenic 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 monovalent Fab format; it is ideal for bioanalytical assay development to measure levels of tocilizumab or biosimilars. It is recommended as capture antibody in PK bridging ELISA with HCA253/HCA253P.
    • Product Type
      Monoclonal Antibody
    • Clone
      AbD21362
    • Isotype
      HuCAL Fab monovalent
    1 Formats Available
      Product CodeApplicationsDatasheetMSDSPack SizeList PriceQuantity
      HCA252E *datasheet pdfdatasheet pdf0.1 mg
      HCA252
      Summary
      Secondary Antibodies
      Negative Isotype Controls
      Useful Reagents
      Positive Controls
      Histology Controls
      More Images
      References
      Reviews
      -
      • Human anti Tocilizumab, clone AbD21362 is a paratope specific, anti-idiotypic antibody that binds to free tocilizumab but not to human Interleukin-6 Receptor (IL-6R) or the drug/IL-6R complex. The antibody can be used to measure the levels of tocilizumab and biosimilar products in bioanalytical assays.

        Clone AbD21362 can be used to develop a pharmacokinetic (PK) bridging assay to measure free drug. This antibody is recommended as the capture antibody, paired with the detection antibody clone AbD21338_hIgG HRP-conjugated and in full immunoglobulin (Ig) format (HCA253).

        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. The therapeutic antibody is directed against IL-6R and 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.

        View a summary of all anti-tocilizumab antibodies
      • Intended Use
      • Product Form
        A monovalent human recombinant Fab 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
      • Preservative Stabilisers
        0.01% Thiomersal
      • Immunogen
        Tocilizumab
      • Purity
      • Affinity
        The monovalent intrinsic affinity of this antibody was measured as KD=0.9 nM by real time, label-free molecular interaction analysis on immobilized tocilizuimab.
      • 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(1)
        (1)
        NB. The Rat anti DYKDDDDK-tag antibody, clone 6F7 is not suitable for use with this antibody as clone 6F7 exhibits cross reactivity with the human IL-6R

      • 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 a capture reagent in a PK bridging ELISA together with HCA253 as the detection reagent.
        Protocol: PK bridging ELISA to measure free drug
      • Immunohistology
      • Histology Positive Control Tissue
      • Immunofluorescence
      • Western Blotting
      • Instructions For Use

      Additional Tocilizumab Antibody Formats

      Formats Clone Applications Sizes available
      Tocilizumab Antibody : Purified AbD21362 E * 0.1 mg
      • Copyright © 2016 Bio-Rad

      Recommended Secondary Antibody

        DescriptionProduct CodePack SizeApplicationsList PriceQuantity
        Mouse anti Penta Histidine Tag:HRPMCA5995P125 µlE, WB
        MCA5995P

        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 TocilizumabHCA2530.1 mgE
            HCA253
            Human anti TocilizumabHCA2540.1 mgE
            HCA254
            Human anti TocilizumabHCA2550.1 mgE
            HCA255
            Human anti TocilizumabHCA2560.1 mgE
            HCA256
            Human anti TocilizumabHCA2570.1 mgE
            HCA257
            Hispec Assay DiluentBUF049A50 mlIY
            BUF049A

            Recommended Positive Controls

              Histology Controls

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