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Multicolor flow cytometric analysis of CD64[a,b] expression on Mouse bone marrow cells. Mouse bone marrow cells were treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes and washed. The cells were preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Rat Anti-CD11b antibody (Cat. No. 553312) and with either BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat. No. 562652; Left Plot) or BD Horizon™ BV605 Mouse Anti-Mouse CD64[a,b] antibody (Cat. No. 569298/569299; Right Plot) at 1 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plot showing the correlated expression of CD64[a,b] (or Ig Isotype control staining) versus CD11b was derived from gated events with the side and forward light-scatter characteristics of viable (DAPI-negative) bone marrow cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
BD Horizon™ BV605 Mouse Anti-Mouse CD64[a,b]
Regulatory Statusの凡例
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
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BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells. However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.
For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant dyes are used in a multicolor flow cytometry panel. Fluorescent dye interactions may cause staining artifacts which may affect data interpretation. The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Although every effort is made to minimize the lot-to-lot variation in the efficiency of the fluorochrome energy transfer, differences in the residual emission from BD Horizon™ BV421 may be observed. Therefore, we recommend that individual compensation controls be performed for every BD Horizon™ BV605 conjugate.
- BD Horizon Brilliant Violet 605 is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,227,187; 8,455,613; 8,575,303; 8,354,239.
- BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- CF™ is a trademark of Biotium, Inc.
- For U.S. patents that may apply, see bd.com/patents.
関連製品
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 (5)
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Bosteels C, Neyt K, Vanheerswynghels M, et al. Inflammatory Type 2 cDCs Acquire Features of cDC1s and Macrophages to Orchestrate Immunity to Respiratory Virus Infection.. Immunity. 2020; 52(6):1039-1056.e9. (Clone-specific: Flow cytometry). View Reference
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Dal-Secco D, Wang J, Zeng Z, et al. A dynamic spectrum of monocytes arising from the in situ reprogramming of CCR2+ monocytes at a site of sterile injury.. J Exp Med. 2015; 212(4):447-56. (Clone-specific: Flow cytometry). View Reference
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Deniset JF, Belke D, Lee WY, et al. Gata6+ Pericardial Cavity Macrophages Relocate to the Injured Heart and Prevent Cardiac Fibrosis.. Immunity. 2019; 51(1):131-140.e5. (Clone-specific: Flow cytometry). View Reference
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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. (Clone-specific). View Reference
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