Preparation And Storage
Recommended Assay Procedures
BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Note: When using high concentrations of antibody, background binding of this dye to erythroid cell subsets (mature erythrocytes and precursors) has been observed. For researchers studying these cell populations, or in cases where light scatter gating does not adequately exclude these cells from the analysis, this background may be an important factor to consider when selecting reagents for panel(s).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For U.S. patents that may apply, see bd.com/patents.
- 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.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
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.
Development References (6)
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Fan Z, Lu Y, Wu X, Mendelsohn J. Antibody-induced epidermal growth factor receptor dimerization mediates inhibition of autocrine proliferation of A431 squamous carcinoma cells.. J Biol Chem. 1994; 269(44):27595-602. (Biology). View Reference
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Goldstein NI, Prewett M, Zuklys K, Rockwell P, Mendelsohn J. Biological efficacy of a chimeric antibody to the epidermal growth factor receptor in a human tumor xenograft model.. Clin Cancer Res. 1995; 1(11):1311-8. (Biology). View Reference
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Matar P, Rojo F, Cassia R, et al. Combined epidermal growth factor receptor targeting with the tyrosine kinase inhibitor gefitinib (ZD1839) and the monoclonal antibody cetuximab (IMC-C225): superiority over single-agent receptor targeting.. Clin Cancer Res. 2004; 10(19):6487-501. (Biology). View Reference
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Patel D, Lahiji A, Patel S, et al. Monoclonal antibody cetuximab binds to and down-regulates constitutively activated epidermal growth factor receptor vIII on the cell surface.. Anticancer Res. 2007; 27(5A):3355-66. (Biology). View Reference
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Sato JD, Kawamoto T, Le AD, Mendelsohn J, Polikoff J, Sato GH. Biological effects in vitro of monoclonal antibodies to human epidermal growth factor receptors.. Mol Biol Med. 1983; 1(5):511-29. (Biology). View Reference
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Wee P, Wang Z. Epidermal Growth Factor Receptor Cell Proliferation Signaling Pathways.. Cancers (Basel). 2017; 9(5):52. (Biology). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.