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PE Rat Anti-Mouse PIR-A/B
PE Rat Anti-Mouse PIR-A/B
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.
Product Details
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BD Pharmingen™
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG1, κ
Recombinant mouse PIR-A1 extracellular domain peptide and the mouse myeloid cell line, WEH13
Flow cytometry (Routinely Tested)
0.2 mg/ml
AB_393628
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.

Recommended Assay Procedures

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.

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. 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.
550349 Rev. 5
Antibody Details
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6C1

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.  

550349 Rev. 5
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
550349 Rev.5
Citations & References
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Development References (6)

  1. Bléry M, Kubagawa H, Chen CC, Vély F, Cooper MD, Vivier E. The paired Ig-like receptor PIR-B is an inhibitory receptor that recruits the protein-tyrosine phosphatase SHP-1. Proc Natl Acad Sci U S A. 1998; 95(5):2446-2451. (Biology). View Reference
  2. Chen CC, Hurez V, Brockenbrough JS, Kubagawa H, Cooper MD. Paternal monoallelic expression of the paired immunoglobulin-like receptors PIR-A and PIR-B. Proc Natl Acad Sci U S A. 1999; 96(12):6868-6872. (Clone-specific). View Reference
  3. Kubagawa H, Burrows PD, Cooper MD. A novel pair of immunoglobulin-like receptors expressed by B cells and myeloid cells. Proc Natl Acad Sci U S A. 1997; 94(10):5261-5266. (Biology). View Reference
  4. Kubagawa H, Chen CC, Ho LH. Biochemical nature and cellular distribution of the paired immunoglobulin-like receptors, PIR-A and PIR-B. J Exp Med. 1999; 189(2):309-318. (Immunogen). View Reference
  5. Maeda A, Kurosaki M, Kurosaki T. Paired immunoglobulin-like receptor (PIR)-A is involved in activating mast cells through its association with Fc receptor gamma chain. J Exp Med. 1998; 188(5):991-995. (Biology). View Reference
  6. Taylor LS, McVicar DW. Functional association of FcepsilonRIgamma with arginine(632) of paired immunoglobulin-like receptor (PIR)-A3 in murine macrophages. Blood. 1999; 94(5):1790-1796. (Biology). View Reference
View All (6) View Less
550349 Rev. 5

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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.