Cetuximab antibody | AbD19376_hIgG1
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 HRP conjugated Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228) at a concentration of 2 μg/ml in HISPEC Assay Diluent (BUF049A) followed by QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated over night with human IgG1/kappa, human IgG1/lambda, rituximab or cetuximab at a concentration of 5 μg/ml. After washing and blocking with PBST+5% BSA, detection was performed using Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228) in a monovalent format, titrated to the given concentrations in PBST+10% human serum, followed by Mouse Anti-Penta Histidine Tag:HRP (MCA5995P) in HISPEC Assay Diluent (BUF049A) and QuantaBlu Fluorogenic Peroxidase Substrate.
A microtiter plate was coated over night with Human Anti-Cetuximab Antibody, clone AbD19834 (HCA220) 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 cetuximab. Detection was performed using HRP conjugated Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228) 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 epidermal growth factor receptor (EGFR) at a concentration of 1 μg/ml. After washing and blocking with PBST+5% BSA, a pre-incubated mixture of cetuximab (CTX, 0.3 μg/ml) plus increasing concentrations of the non-inhibitory Human Anti-Cetuximab Antibody (HCA228) was added. In parallel experiments, HCA228 was replaced with inhibitory Human Anti-Cetuximab Antibodies of varying affinity (HCA220, HCA221 or HCA223). For this assay, HCA221, HCA223 and HCA228 were used in their monovalent Fab format. Free cetuximab, still capable of binding to the EGFR-coated plate, was detected using Goat Anti-Human IgG F(ab')2:HRP (STAR126P).
Human anti Cetuximab:HRP
Anti-Cetuximab Antibody is a recombinant, non-inhibitory, anti-idiotypic antibody for use in bioanalytical assays for cetuximab and biosimilars. It is labeled with HRP, and is recommended as the detection antibody in a PK bridging ELISA with HCA220 to measure total drug.
Human anti Cetuximab
Anti-Cetuximab Antibody is a non-inhibitory anti-idiotypic IgG1 antibody; it is ideal for bioanalytical assays for cetuximab and biosimilars. It does not inhibit the binding of cetuximab to EGFR; it is suitable as a detection antibody in PK bridging ELISA with HCA220 to measure total drug.
- Product Type
- Monoclonal Antibody
- Clone
- AbD19376_hIgG1
- Isotype
- IgG1
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA228P | E | 0.1 mg |
|
|
| HCA228 | E | 0.1 mg |
|
Product Details
- Product Form
- Human IgG1 antibody selected from the HuCAL® phage display library and expressed in a human cell line. Conjugated to horseradish peroxidase (HRP) - liquid.
- 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.
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Preparation
- Purified IgG prepared by affinity chromatography on Protein A
- Buffer Solution
- Phosphate buffered saline
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Preservative Stabilisers
- 0.01% Thiomersal
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD=39 nM by real time, label-free molecular interaction analysis on immobilized cetuximab.
- Approx. Protein Concentrations
- IgG concentration 0.1 mg/ml
- Approx. Protein Concentrations
- Antibody concentration 0.5 mg/ml
Storage Information
- Storage
- Store at -70oC.
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. - 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. - Guarantee
- 12 months from date of despatch
- Guarantee
- 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.
- 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.
- 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 Cetuximab antibody
| Application Name | Verified | Min Dilution | Max Dilution |
|---|---|---|---|
| ELISA | |||
| 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.
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
- 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 antibody in a cetuximab bridging ELISA for PK assay development together with HCA220 as the capture reagent.
Protocol: PK bridging ELISA to measure total drug - ELISA
- This product may be used in a direct ELISA or when conjugated to HRP, as a detection antibody in a cetuximab bridging ELISA for PK assay development together with HCA220 as the capture reagent.
Protocol: PK bridging ELISA to measure total drug
Copyright © 2021 Bio-Rad Antibodies (formerly AbD Serotec)
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Cetuximab | HCA220 | E | 0.1 mg |
|
|
| Human anti Cetuximab | HCA221 | E | 0.1 mg |
|
|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Cetuximab | HCA220 | E | 0.1 mg |
|
|
| Human anti Cetuximab | HCA221 | E | 0.1 mg |
|
|
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody |
Product Specific References
References for Cetuximab antibody
-
Bose, M.E. et al. (2016) Simulated Respiratory Secretion for Use in the Development of Influenza Diagnostic Assays.
PLoS One. 11 (11): e0166800. -
Kashiwagi, N. et al. (2017) Method for measuring anti-drug antibody
US Patent Application US20170315118A1
Further Reading
-
Gomez, D. et al. (2013) Cetuximab therapy in the treatment of metastatic colorectal cancer: the future frontier?
Int J Surg. 11 (7): 507-13.
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