Description
The Cetuximab297.rMAb is a research grade, chimeric recombinant human IgG1, kappa antibody that specifically recognizes the extracellular domain of Epidermal Growth Factor Receptor (EGFR) which is also known as HER1 or ErbB-1. The Cetuximab.rMAb is a biosimilar antibody that uses the same variable region sequences as the chimeric (mouse/human) therapeutic antibody, Cetuximab, combined with constant region sequences derived from a consensus sequence of human IgG1, kappa. The asparagine at position 297 of the constant heavy chain has been replaced with alanine (N297A) to further reduce Fc receptor interactions of this reagent. Cetuximab was derived from the Mouse 225 antibody that specifically binds to the extracellular domain of human EGFR and blocks its signaling activity. EGFR is a ~170 kDa single pass type I transmembrane glycoprotein that is encoded by EGFR which belongs to the HER/ErbB family of receptor tyrosine kinases (RTKs) that includes HER2 (ErbB-2, Neu, CD340), HER3 (ErbB-3), and HER4 (ErbB-4). EGFR is expressed on various cells including epithelial and endothelial cells. It plays and important role in their proliferation, survival, and motility in response to various ligands including EGF, heparin-binding EGF, amphiregulin, betacellulin, epiregulin, vaccinia virus growth factor and TGF-α. Upon ligand binding, EGFR undergoes homodimerization or heterodimerization with the other ErbB family members. This induces EGFR transautophosphorylation and stimulation of numerous downstream intracellular signaling pathways including the RAS/RAF/MEK/ERK, PLC-γ/PKC, PI3K/AKT, JAK/STAT, and NF-κB pathways. Activated EGFR-ligand complexes are removed from the cell surface via endocytosis and are degraded in the lysosomes to attenuate the signaling. Aberrant EGFR expression and signaling has been reported in inflammatory and malignant diseases. The Cetuximab297.rMAb is intended for research use only. It is not intended for use in therapeutic or diagnostic procedures for humans or animals.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions that minimize unconjugated dye and antibody.
1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
2. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
3. For U.S. patents that may apply, see bd.com/patents.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
6. The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
7. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
9. An isotype control should be used at the same concentration as the antibody of interest.
10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
11. Tandem fluorochromes contain both an energy donor and an energy acceptor. Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
