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Two-color analysis of the expression of PIR-A/B on mouse spleen cells. BALB/c splenocytes were preincubated with Mouse BD Fc Block™ (Cat. no. 553141/553142) and simultaneously stained with PE- conjugated 6C1 (both panels) and either FITCconjugated 1D3 (anti-mouse CD19, Cat. no. 557398/553785, left panel) or FITC- conjugated M1/70 (anti-mouse CD11b, Cat. no. 557396/553310, right panel) monoclonal antibodies. Left panel demonstrates that the majority of B lymphocytes (CD19-positive cells) express intermediate levels of PIR- A/B, while the right panel shows that most myeloid cells (CD11b- or Mac-1-positive) express high levels of PIR-A/B. Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The quadrant positions are based upon analyses of isotype controls.
Two-color analysis of the expression of PIR-A/B on mouse spleen cells. BALB/c splenocytes were preincubated with Mouse BD Fc Block™ (Cat. no. 553141/553142) and simultaneously stained with PE- conjugated 6C1 (both panels) and either FITCconjugated 1D3 (anti-mouse CD19, Cat. no. 557398/553785, left panel) or FITC- conjugated M1/70 (anti-mouse CD11b, Cat. no. 557396/553310, right panel) monoclonal antibodies. Left panel demonstrates that the majority of B lymphocytes (CD19-positive cells) express intermediate levels of PIR- A/B, while the right panel shows that most myeloid cells (CD11b- or Mac-1-positive) express high levels of PIR-A/B. Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The quadrant positions are based upon analyses of isotype controls.
Two-color analysis of the expression of PIR-A/B on mouse spleen cells. BALB/c splenocytes were preincubated with Mouse BD Fc Block™ (Cat. no. 553141/553142) and simultaneously stained with PE- conjugated 6C1 (both panels) and either FITCconjugated 1D3 (anti-mouse CD19, Cat. no. 557398/553785, left panel) or FITC- conjugated M1/70 (anti-mouse CD11b, Cat. no. 557396/553310, right panel) monoclonal antibodies. Left panel demonstrates that the majority of B lymphocytes (CD19-positive cells) express intermediate levels of PIR- A/B, while the right panel shows that most myeloid cells (CD11b- or Mac-1-positive) express high levels of PIR-A/B. Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The quadrant positions are based upon analyses of isotype controls.
Two-color analysis of the expression of PIR-A/B on mouse spleen cells. BALB/c splenocytes were preincubated with Mouse BD Fc Block™ (Cat. no. 553141/553142) and simultaneously stained with PE- conjugated 6C1 (both panels) and either FITCconjugated 1D3 (anti-mouse CD19, Cat. no. 557398/553785, left panel) or FITC- conjugated M1/70 (anti-mouse CD11b, Cat. no. 557396/553310, right panel) monoclonal antibodies. Left panel demonstrates that the majority of B lymphocytes (CD19-positive cells) express intermediate levels of PIR- A/B, while the right panel shows that most myeloid cells (CD11b- or Mac-1-positive) express high levels of PIR-A/B. Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The quadrant positions are based upon analyses of isotype controls.
Two-color analysis of the expression of PIR-A/B on mouse spleen cells. BALB/c splenocytes were preincubated with Mouse BD Fc Block™ (Cat. no. 553141/553142) and simultaneously stained with PE- conjugated 6C1 (both panels) and either FITCconjugated 1D3 (anti-mouse CD19, Cat. no. 557398/553785, left panel) or FITC- conjugated M1/70 (anti-mouse CD11b, Cat. no. 557396/553310, right panel) monoclonal antibodies. Left panel demonstrates that the majority of B lymphocytes (CD19-positive cells) express intermediate levels of PIR- A/B, while the right panel shows that most myeloid cells (CD11b- or Mac-1-positive) express high levels of PIR-A/B. Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The quadrant positions are based upon analyses of isotype controls.
Two-color analysis of the expression of PIR-A/B on mouse spleen cells. BALB/c splenocytes were preincubated with Mouse BD Fc Block™ (Cat. no. 553141/553142) and simultaneously stained with PE- conjugated 6C1 (both panels) and either FITCconjugated 1D3 (anti-mouse CD19, Cat. no. 557398/553785, left panel) or FITC- conjugated M1/70 (anti-mouse CD11b, Cat. no. 557396/553310, right panel) monoclonal antibodies. Left panel demonstrates that the majority of B lymphocytes (CD19-positive cells) express intermediate levels of PIR- A/B, while the right panel shows that most myeloid cells (CD11b- or Mac-1-positive) express high levels of PIR-A/B. Flow cytometry was performed on a BD FACScan™ Flow Cytometry System. The quadrant positions are based upon analyses of isotype controls.
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This antibody has been tested by immunofluorescent staining (≤1 µg/million cells) with flow cytometric analysis to assure specificity and reactivity. Mouse BD Fc Block. (Cat. no. 553141/553142) should be used when staining with 6C1 antibody. Since applications vary, each investigator must determine dilutions appropriate for individual use.
The 6C1 monoclonal antibody specifically recognizes the common epitopes of PIR-A and PIR-B (Paired Immunoglobulin-liked Receptors) in all mouse strains tested (A/J, BALB/cJ, C3H, C57BL/6, DBA/1, DBA/2, NZB, and SJL). PIR-A and PIR-B are type-I transmembrane glycoproteins containing six Ig-like domains. There are multiple PIR-A proteins which are activating receptors by virtue of their intracellular association with the ITAM (Immunoreceptor Tyrosine-based Activation Motif)- containing Fc receptor γ chain (FcRγ) in mast cells and macrophages. FcRγ expression is required for PIR-A cell-surface expression on dendritic cells, B cells, and myeloid lineages. In contrast, PIR-B is an inhibitory receptor which contains various ITIMs (Immunoreceptor Tyrosine-based Inhibitory Motifs) in its cytoplasmic domain. These receptors are expressed on B cells, granulocytes, mast cells, dendritic cells, and monocytes/macrophages, but not on thymocytes, T lymphocytes, erythroid lineage cells, or NK cells. The level of cell-surface expression of these receptors increases as a function of B-cell activation and myeloid- and B-lineage differentiation. The 6C1 antibody immunoprecipitates molecules of 85- and 125-kDa, which correspond to PIRA and PIR-B, respectively.
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