Tocilizumab antibody | AbD21362
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
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
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 ®
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 ®
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 ®
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 ®
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 ®
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
Human anti Tocilizumab
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
| Product Code | Applications | Datasheet | MSDS | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|---|
| HCA252 | E* | 0.1 mg |
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
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
Product Details
- 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.
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Immunogen
- Tocilizumab
- 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
Storage Information
- 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
More Information
- 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.
Actemra® and RoActemra® are registered trademarks of Hoffman-La Roche and Chugai Pharmaceuticals co., Ltd.
His-tag is a registered trademark of EMD Biosciences. - 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
Applications of Tocilizumab antibody
| Application Name | Verified | Min Dilution | Max 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
- 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
Copyright © 2018 Bio-Rad Antibodies (formerly AbD Serotec)
Secondary Antibodies Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | 125 µl | E WB |
Useful Reagents Available
| Description | Product Code | Pack Size | Applications | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | 50 ml | E IY | ||
| Human anti Tocilizumab | HCA253 | 0.1 mg | E | ||
| Human anti Tocilizumab | HCA254 | 0.1 mg | E | ||
| Human anti Tocilizumab | HCA255 | 0.1 mg | E | ||
| Human anti Tocilizumab | HCA256 | 0.1 mg | E | ||
| Human anti Tocilizumab | HCA257 | 0.1 mg | E | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK001P | 1 Conjugation For 400µg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | 3 Conjugations For 400µg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK003P | 1 Conjugation For 4mg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK004P | 5 Conjugations For 4mg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK005P | 1 Conjugation For 20mg Antibody | CJ | ||
| LYNX Rapid HRP Antibody Conjugation Kit | LNK006P | 3 Conjugations For 40µg Antibody | CJ |
