Mouse monoclonal antibody is produced from a hybridoma resulting from fusion of SP2/0 myeloma and B-lymphocytes obtained from a mouse immunized with Cetuximab F(ab')2.
Recognizes Cetuximab specifically, no cross reactivity with other humanized antibodies.
The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with biotins using standard chemical labeling method. A specially optimized long-spacer biotin reagent (13.5 angstroms) is used in this product to minimize potential steric hindrance.
The biotin to protein ratio is 1-3 as determined by the HABA assay.
>90% as determined by SDS-PAGE.
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4. Normally trehalose is added as protectant before lyophilization.
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
No activity loss was observed after storage at:
- 4-8°C for 12 months in lyophilized state;
- -70°C for 3 years under sterile conditions after reconstitution.
Biotinylated Anti-Cetuximab Antibodies (recommended for PK/PD) on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
Detection of cetuximab by bridging ELISA in serum. Immobilized Anti-Cetuximab Antibodies (Cat. No. CEB-Y29) at 2 μg/ml, added increasing concentrations of cetuximab (10% human serum) and then added Biotinylated Anti-Cetuximab Antibodies (Cat. No. CEB-BY31) at 1 μg/ml. Detection was performed using HRP-conjugated streptavidin with a sensitivity of 6 ng/ml.
Immobilized cetuximab at 0.1 μg/mL (100 μL/well) can bind Biotinylated Anti-Cetuximab Antibodies (Cat. No. CEB-BY31) with a linear range of 0.39-25 ng/mL.
ELISA analysis shows that the binding of cetuximab to Human EGF R, His Tag (Cat. No. EGR-H5222) was inhibited by increasing concentration of Biotinylated Anti-Cetuximab Antibodies (Cat. No. CEB-BY31). The concentration of cetuximab used is 3 ng/ml.
Cetuximab is an epidermal growth factor receptor binding FAB. Cetuximab is composed of the Fv (variable; antigen-binding) regions of the 225 murine EGFr monoclonal antibody specific for the N-terminal portion of human EGFr with human IgG1 heavy and kappa light chain constant (framework) regions.