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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)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.
BD Pharmingen™ Alexa Fluor® 647 Mouse anti-Mouse CD64 a and b Alloantigens
Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
- 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.
- Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
- An isotype control should be used at the same concentration as the antibody of interest.
Companion Products
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.
Development References (2)
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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.
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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
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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.