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PE Mouse anti-Mouse CD64 a and b Alloantigens
PE Mouse anti-Mouse CD64 a and b Alloantigens

Flow cytometric analysis of PE-conjugated anti-mouse CD64 recognizing a and b Alloantigens on mouse bone marrow cells. Isolated murine bone marrow cells were preincubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142).  The cells were then stained with FITC anti-CD11b (clone M1/70, catalog number No. 553310) and either PE anti-CD64 (clone X54-5/7.1, Cat. No. 558455, right panel) or a PE mouset IgG1 isotype control (catalog number 550617, left panel).  Flow cytometry was performed on a BD FACSCalibur™ System and the dot plots were derived from the gated events based on light scattering characteristics of viable bone marrow cells.

Flow cytometric analysis of PE-conjugated anti-mouse CD64 recognizing a and b Alloantigens on mouse bone marrow cells. Isolated murine bone marrow cells were preincubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142).  The cells were then stained with FITC anti-CD11b (clone M1/70, catalog number No. 553310) and either PE anti-CD64 (clone X54-5/7.1, Cat. No. 558455, right panel) or a PE mouset IgG1 isotype control (catalog number 550617, left panel).  Flow cytometry was performed on a BD FACSCalibur™ System and the dot plots were derived from the gated events based on light scattering characteristics of viable bone marrow cells.

Product Details
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BD Pharmingen™
Fcgr1; FcRI; Fcg1; Fc-gamma RI; Fc receptor IgG high affinity I; IGGHAFC
Mouse (QC Testing)
Mouse NOD/Lt IgG1, κ
Mouse CD64 a Alloantigen
Flow cytometry (Routinely Tested)
0.2 mg/ml
14129
AB_647241
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

For flow cytometry of cell suspensions from peripheral lymphoid tissues, it is recommended that the cells be pre-incubated with Mouse BD Fc Block™ purified anti-mouse CD16/CD32 mAb 2.4G2 (Cat. no.553141/553142).

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.
558455 Rev. 1
Antibody Details
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X54-5/7.1

The X54-5/7.1 monoclonal antibody specifically recognizes FcγRI (CD64) encoded by the more common Fcgr1a and Fcgr1b alleles.  The alloantigens generated by the Fcgr1a and Fcgr1b alleles, have been confirmed positive in mouse strains BALB/c and C57BL/6 and reported positive in strains 129, A, AKR, ALR, BUB, C3H, C57BL/10, C57BLKS, C57BR, C58, CBA, CE, DBA/2, HRS, MRL, NON, NZB, NZO, NZW, PL, SJL, ST, SWR.  The a and b alloantigens have been reported negative in mouse strains ABH, NOD.  CD64, a key receptor in the development of immune responses, has a dual role as a low affinity receptor for IgG3 and high affinity receptor for IgG2a linking innate and adaptive immunities.  CD64 mediates endocytosis, phagocytosis, antibody-dependent cellular toxicity, cytokine release and superoxide generation.  CD64 is expressed largely on macrophages and dendritic cells.  For more information regarding clone X54-5/7.1 and the alloantigens it recognizes, please refer to the reference by Tan et al listed below. 

558455 Rev. 1
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
558455 Rev.1
Citations & References
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Development References (2)

  1. Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997.
  2. Tan PS, Gavin AL, Barnes N, et al. Unique monoclonal antibodies define expression of Fc gamma RI on macrophages and mast cell lines and demonstrate heterogeneity among subcutaneous and other dendritic cells. J Immunol. 2003; 170(5):2549-2556. (Clone-specific). View Reference
558455 Rev. 1

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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.