Tocilizumab antibody | AbD22155_hIgG1
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 AbD22155_hIgG1) at a concentration of 2 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate.
A microtiter plate was coated over night with tocilizumab, human interleukin-6 receptor (hIL-6R), human IgG1/kappa or human IgG1/lambda at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, the hIL-6R/tocilizumab complex was formed by adding tocilizumab to hIL-6R-coated wells for an incubation period of 1 hour. Detection was subsequently performed using HRP-conjugated Human anti tocilizumab (clone AbD22155_hIgG1) titrated to the given concentrations in PBST, followed by QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements.
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 AbD22155_hIgG1) at a concentration of 1 μg/ml in HISPEC assay diluent (BUF049A) and QuantaBlu fluorogenic peroxidase substrate. Data is presented as the mean of three measurements.
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 non-inhibitory Human anti tocilizumab clone AbD22155_hIgG1 or the inhibitory clone AbD21338_hIgG (HCA253) 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.
Human anti Tocilizumab:HRP
- Product Type
- Monoclonal Antibody
- Clone
- AbD22155_hIgG1
- Isotype
- IgG1
- Specificity
- Tocilizumab
Antibody Quality and Performance Guarantee
| Human Anti-Tocilizumab Antibody, clone AbD22155_hIgG1 is a non-paratope specific, anti-idiotypic antibody that binds to free tocilizumab and also to tocilizumab bound to its target, human interleukin-6 receptor (IL-6R). The non-inhibitory antibody can be used to measure total levels of tocilizumab and biosimilar products in bioanalytical assays. Clone AbD22155_hIgG1 can be used to develop a pharmacokinetic (PK) bridging assay to measure total drug levels (free drug, partially bound and fully bound). This non-inhibitory antibody is recommended as the detection antibody, paired with the inhibitory capture antibody clone AbD21362 (HCA252). This combination enables measurement of free and partially bound tocilizumab, which is equivalent to total drug when trough serum levels are above 10 μg/ml. Tocilizumab (RoActemra/Actemra) 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 |
- Product Form
- Human IgG1 antibody (lambda light chain) selected from the HuCAL phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) - liquid.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Source
- HKB-11
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Tocilizumab and tocilizumab-sIL-6R complex
- Affinity
- The intrinsic affinity of the monovalent form of this antibody is KD = 31 nM as measured by real time, label free molecular interaction analysis on immobilized tocilizumab.
- Approx. Protein Concentrations
- IgG concentration 1.0 mg/ml
- Regulatory
- For research purposes only
- Guarantee
- 12 months from date of despatch
- Acknowledgements
- This product and/or its use is covered by claims of U.S. patents, and/or pending U.S. and non-U.S. patent applications owned by or under license to Bio-Rad Laboratories, Inc. See bio-rad.com/en-us/trademarks for details.
Actemra and RoActemra are registered trademarks of Hoffman-La Roche and Chugai Pharmaceuticals co., Ltd.
This product is shipped frozen.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
When ready to use, thaw and aliquot the sample as needed. Store aliquots at -70°C, if available, otherwise store at -20°C. It is not recommended to keep aliquots at 4°C for more than one week.
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 Name | Verified | Min Dilution | Max 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.
- 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 total drug.
| Description | Product Code | Applications | Pack Size | List Price | Your Price | Quantity | |
|---|---|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | Log in | |||
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| Description | Hispec Assay Diluent | ||||||
| Human anti Tocilizumab | HCA252 | E * | 0.1 mg |
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| Description | Human anti Tocilizumab | ||||||
| Human anti Tocilizumab | HCA253 | E | 0.1 mg |
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| Description | Human anti Tocilizumab | ||||||
| Human anti Tocilizumab | HCA254 | E | 0.1 mg |
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| Description | Human anti Tocilizumab | ||||||
| Human anti Tocilizumab | HCA255 | E | 0.1 mg |
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| Description | Human anti Tocilizumab | ||||||
| Human anti Tocilizumab | HCA256 | E | 0.1 mg |
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| Description | Human anti Tocilizumab | ||||||
References for Tocilizumab antibody
-
Kashiwagi, N. et al. (2017) Method for measuring anti-drug antibody
US Patent Application US20170315118A1
- Synonyms
- ACTEMRA®
- ROACTEMRA®
- 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.
HCA257P
153184
159732
162312
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