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Alexa Fluor® 647 Mouse anti-Mouse CD64 a and b Alloantigens

BD Pharmingen™ Alexa Fluor® 647 Mouse anti-Mouse CD64 a and b Alloantigens

Clone X54-5/7.1 (also known as X54-5/7.1)

(RUO)
Alexa Fluor® 647 Mouse anti-Mouse CD64 a and b Alloantigens
Flow cytometric analysis of Alexa Fluor® 647 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, Cat. No. 553310) and either Alexa Fluor® 647 Anti-CD64 (clone X54-5/7.1, Cat. No. 558539, right panel) or a Alexa Fluor® 647 Mouse IgG1 isotype control (Cat. No. 557732, 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 Alexa Fluor® 647 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, Cat. No. 553310) and either Alexa Fluor® 647 Anti-CD64 (clone X54-5/7.1, Cat. No. 558539, right panel) or a Alexa Fluor® 647 Mouse IgG1 isotype control (Cat. No. 557732, 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_647120
Aqueous buffered solution containing protein stabilizer and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  4. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  5. 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.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. An isotype control should be used at the same concentration as the antibody of interest.
558539 Rev. 2
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. 

558539 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
558539 Rev.2
Citations & References
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View product citations for antibody "558539" on CiteAb

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. (Biology). View Reference
558539 Rev. 2

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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.