Cetuximab antibody | AbD19834
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-Cetuximab Antibody, clone AbD19834 (HCA220) at a concentration of 2 μg/ml, 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 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 AbD19834 (HCA220) 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 1 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 AbD19830_hIgG1 (HCA221P) 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 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 the non-inhibitory Human Anti-Cetuximab Antibody, clone AbD19376_hIgG1 (HCA228P) 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.
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 Human Anti-Cetuximab Antibody HCA220, HCA221 or HCA223, or the non-inhibitory antibody HCA228, was added. For this assay HCA221, HCA223 and HCA228 were used in the 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
Anti-Cetuximab Antibody is a recombinant, inhibitory, anti-idiotypic antibody in monovalent Fab format; it is ideal for bioanalytical assay development for cetuximab and biosimilars. It is suitable as a capture antibody in PK bridging ELISA format with HCA221 for an assay to detect free drug.
- Product Type
- Monoclonal Antibody
- Clone
- AbD19834
- Isotype
- HuCAL Fab monovalent
| Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|
| HCA220 | E | 0.1 mg |
|
Product Details
- Product Form
- A monovalent human recombinant Fab (lambda 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.
- Preparation
- Metal chelate affinity chromatography
- Buffer Solution
- Phosphate buffered saline
- Preservative Stabilisers
- 0.01% Thiomersal
- Affinity
- The monovalent intrinsic affinity of this antibody was measured as KD = 0.5 nM by real time, label free molecular interaction analysis on immobilized cetuximab.
- Approx. Protein Concentrations
- Total protein 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. - 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.
Erbitux® is a registered trademark of ImClone LLC, a wholly-owned subsidiary of Eli Lilly and Company.
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 Cetuximab antibody
| 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 coating antibody in PK bridging assays together with HRP-conjugated HCA221 or HCA228 as the detection reagent.
Protocol: PK bridging ELISA to measure free drug
Copyright © 2021 Bio-Rad Antibodies (formerly AbD Serotec)
Secondary Antibodies Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Rabbit anti DYKDDDDK Tag:HRP | AHP1074P | E WB | 0.1 mg |
|
|
| Goat anti Histidine Tag:Biotin | AHP1656B | E WB | 0.1 mg |
|
|
| Mouse anti Histidine Tag:Alk.Phos. | MCA1396A | E WB | 0.1 mg |
|
|
| Mouse anti Histidine Tag:Biotin | MCA1396B | E WB | 0.1 mg |
|
|
| Rat anti DYKDDDDK Tag:HRP | MCA4764P | E P WB | 0.1 mg |
|
|
| Mouse anti Penta Histidine Tag:HRP | MCA5995P | E WB | 125 µl |
|
|
| Goat anti Human IgG F(ab')2:HRP | STAR126P | E WB | 0.5 ml |
|
Useful Reagents Available
| Description | Product Code | Applications | Pack Size | List Price | Quantity |
|---|---|---|---|---|---|
| Hispec Assay Diluent | BUF049A | E IY | 50 ml | ||
| Human anti Cetuximab | HCA221 | E | 0.1 mg |
|
|
| Human anti Cetuximab | HCA228 | E | 0.1 mg |
|
|
| LYNX Rapid HRP Antibody Conjugation Kit | LNK002P | CJ | 3 Conjugations For 400µg Antibody | ||
| Mouse anti Human IgG (Fc) CH2 Domain:HRP | MCA647P | C* E | 0.2 mg |
|
Product Specific References
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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