Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Mouse CD66a (CEACAM1a) antibody (Cat. No. 750885), right panel) and Isotype Control (left panel) on live BALB/c mouse splenocytes. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.
Multiparameter flow cytometric analysis using BD OptiBuild™ BV421 Mouse Anti-Mouse CD66a (CEACAM1a) antibody (Cat. No. 750885), right panel) and Isotype Control (left panel) on live BALB/c mouse splenocytes. Flow cytometry was performed using a BD LSRFortessa™ Flow Cytometer System.
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 CompBead 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 two or more BD Horizon Brilliant dyes are used in the same experiment. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. 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).
Product Notices
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- BD Horizon Brilliant Violet 421 is covered by one or more of the following US patents: 8,158,444; 8,362,193; 8,575,303; 8,354,239.
Companion Products
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.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.
Development References (3)
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Dveksler GS, Dieffenbach CW, Cardellichio CB, et al. J Virol. 1993; 67(1):1-8. (Clone-specific). View Reference
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Dveksler GS, Pensiero MN, Cardellichio CB, et al. Cloning of the mouse hepatitis virus (MHV) receptor: expression in human and hamster cell lines confers susceptibility to MHV. J Virol. 1991; 65(12):6881-6891. (Clone-specific). View Reference
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Williams RK, Jiang GS, Snyder SW, Frana MF, Holmes KV.. Purification of the 110-kilodalton glycoprotein receptor for mouse hepatitis virus (MHV)-A59 from mouse liver and identification of a nonfunctional, homologous protein in MHV-resistant SJL/J mice. J Virol. 1990; 64(8):3817-3823. (Immunogen). View Reference
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