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Multicolor flow cytometric analysis of CD66a (CEACAM1a) expression on viable mouse splenic leucocytes. BALB/c mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 561880/553092) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-CD66a (CEACAM1a) antibody (Cat. No. 566828; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD66a (CEACAM1) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD66a (CEACAM1a) expression on viable mouse splenic leucocytes. BALB/c mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 561880/553092) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-CD66a (CEACAM1a) antibody (Cat. No. 566828; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD66a (CEACAM1) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD66a (CEACAM1a) expression on viable mouse splenic leucocytes. BALB/c mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 561880/553092) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-CD66a (CEACAM1a) antibody (Cat. No. 566828; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD66a (CEACAM1) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD66a (CEACAM1a) expression on viable mouse splenic leucocytes. BALB/c mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 561880/553092) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-CD66a (CEACAM1a) antibody (Cat. No. 566828; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD66a (CEACAM1) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD66a (CEACAM1a) expression on viable mouse splenic leucocytes. BALB/c mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 561880/553092) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-CD66a (CEACAM1a) antibody (Cat. No. 566828; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD66a (CEACAM1) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD66a (CEACAM1a) expression on viable mouse splenic leucocytes. BALB/c mouse spleen cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes, washed, and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) (Cat. No. 553141/553142). The cells were then stained with APC Rat Anti-Mouse CD45R/B220 antibody (Cat. No. 561880/553092) and either PE Mouse IgG1, κ Isotype Control (Cat. No. 554680; Left Plot) or PE Mouse Anti-CD66a (CEACAM1a) antibody (Cat. No. 566828; Right Plot) at 0.25 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The pseudocolor density plot showing the correlated expression of CD66a (CEACAM1) [or Ig Isotype control staining] versus CD45R/B220 was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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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.
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