Description
The CC1 monoclonal antibody specifically recognizes carcinoembryonic antigen-related cell adhesion molecule 1a (CEACAM1a or CEACAM1[a]), an allotypic form of CEACAM1 which is also known as CD66a, Murine hepatitis virus receptor (MHV-R), or Biliary glycoprotein 1 (BGP-1). Four known isoforms of mouse CD66a arise from alternative splicing of RNA transcripts encoded by Ceacam1, a member of the carcinoembryonic antigen (CEA) family and Ig gene superfamily. These isoforms are type I transmembrane proteins that include a heavily glycosylated extracellular region with an N-terminal IgV-like domain and up to three IgC2-like domains followed by a transmembrane region and a cytoplasmic tail of relatively short (10 amino acids) or long (73 amino acids) length. The cytoplasmic tails enable interactions with other intracellular molecules to initiate or regulate cellular responses. The two CD66a isoforms with long cytoplasmic tails contain immunoreceptor tyrosine-based inhibitory motifs (ITIMs) that may enable CD66a to function as an immune checkpoint inhibitor. CD66a is expressed on a variety of cell types including certain epithelial cells, endothelial cells, B cells, activated T cells, NK cells, monocytes, dendritic cells (DC), and neutrophils. CD66a (CEACAM1a) is a multifunctional protein. Through their IgV-like domain, CD66a (CEACAM1a) molecules function as homophilic and heterophilic intercellular adhesion molecules. They can also function as MHV-Rs, angiogenic factors, regulators of cellular proliferation and differentiation, and tumor suppressors. Two distinct Ceacam1 alleles, (a and b), exist because of differences in their IgV-like domain gene sequences. Ceacam1a is found in most inbred mouse strains including BALB/c, C57BL/6, and C3H mice whereas Ceacam1b is found in SJL mice. CD66a (CEACAM1a) proteins are specifically bound by the CC1 antibody whereas CEACAM1b proteins are not. The CC1 antibody recognizes an epitope in the N-terminal domain of mouse CD66a (CEACAM1a) proteins.
Bead-based compensation or unmixing controls, such as BD® CompBeads or BD™ SpectraComp™, can be used as surrogates to assess fluorescence spillover when bound to fluorochrome-conjugated antibodies. Although these beads have spectral properties similar to cells, variations in spectral emission may occur, resulting in differing spillover values compared to biological controls. Therefore, it is considered best practice to compare the spillover obtained from cells and bead-based controls when using BD® CompBeads or BD™ SpectraComp™ for the first time, to ensure they are appropriate for the intended application.
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. Alexa Fluor™ is a trademark of Life Technologies Corporation.
