BD Horizon™ RY703 Mouse Anti-Human CD64
Clone 10.1 (RUO)
Multiparameter flow cytometric analysis of CD64 expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564220), and stained with either BD Horizon™ RY703 Mouse IgG1, κ Isotype Control (Cat. No. 571453; Left Plot) or BD Horizon™ RY703 Mouse Anti-Human CD64 antibody (Cat. No. 571787/571838; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD64 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a 5-laser Flow Cytometer System and FlowJo™ Software.
Multiparameter flow cytometric analysis of CD64 expression on Human peripheral blood leukocyte populations. Human whole blood was treated with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899) to lyse erythrocytes. The leukocytes were washed, preincubated with BD Pharmingen™ Human BD Fc Block™ (Cat. No. 564220), and stained with either BD Horizon™ RY703 Mouse IgG1, κ Isotype Control (Cat. No. 571453; Left Plot) or BD Horizon™ RY703 Mouse Anti-Human CD64 antibody (Cat. No. 571787/571838; Right Plot). The bivariate pseudocolor density plot showing the correlated expression of CD64 (or Ig Isotype control staining) versus side light-scatter (SSC-A) signals was derived from gated events with the forward and side light-scatter characteristics of viable leukocytes. Flow cytometry and data analysis were performed using a 5-laser Flow Cytometer System and FlowJo™ Software.
Product Details
BD Horizon™
FCGR1; FcRI; Fc-gamma RI; IgG Fc Receptor I; High affinity IgG FcR1
Human (QC Testing)
Mouse BALB/c IgG1, κ
Human Rheumatoid synovial fluid cells and fibronectin-purified monocytes
Flow cytometry (Routinely Tested)
5 µl/test
VI MA36
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 to the dye under optimum conditions and unconjugated antibody and free dye were removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Recommended Assay Procedures
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.
Product Notices
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
- 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.
- This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- An isotype control should be used at the same concentration as the antibody of interest.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
- Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
- Tandem fluorochromes contain both an energy donor and an energy acceptor. 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 the donor may be observed. Additionally, multi-laser cytometers may directly excite both the donor and acceptor fluorochromes. Therefore, we recommend for every tandem conjugate, a matched individual single-stain control be acquired for generating a compensation or spectral unmixing matrix.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- For U.S. patents that may apply, see bd.com/patents.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
RY703 Mouse IgG1, κ Isotype Control RUO
Size
0.1 mg
Cat No.
571453
Lysing Buffer RUO
Size
100 mL
Cat No.
555899
Lysing Solution 10X Concentrate IVD
Size
100 mL
Cat No.
349202
Human BD Fc Block™ RUO
Size
0.25 mg
Cat No.
564220
571787 Rev. 1
Antibody Details
10.1
The 10.1 monoclonal antibody specifically binds to CD64, a 72 kDa type I transmembrane glycoprotein that is a high affinity receptor for human IgG (FcγRI), especially the IgG1 and IgG3 subclasses. CD64 is expressed on monocytes, macrophages, dendritic cells, granulocytes activated with interferon-gamma and early myeloid lineage cells. CD64 associates with a signaling FcRγ homodimer to form the functional high affinity FcγRI complex. CD64 functions in both innate and adaptive immune responses and mediates endocytosis, phagocytosis, antigen presentation, antibody-dependent cellular toxicity, cytokine release and superoxide generation.
571787 Rev. 1
Format Details
RY703
The BD Horizon RealYellow™ 703 (RY703) Dye is part of the BD® family of yellow-green dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 557-nm and an emission maximum (Em Max) at 703-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RY703 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY703 can be used as an alternative to PE-Cy5.5 and we recommend using an optical filter centered near 700-nm (eg, a 695/40-nm bandpass filter).
RY703
Yellow-Green 561 nm
557 nm
703 nm
571787 Rev.1
Citations & References
View product citations for antibody "571787" on CiteAb
Development References (6)
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CD64. In: Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007:151.
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Dougherty GJ, Selvendran Y, Murdoch S, Palmer DG, Hogg N. The human mononuclear phagocyte high-affinity Fc receptor, FcRI, defined by a monoclonal antibody, 10.1. Eur J Immunol. 1987; 17(10):1453-1459. (Immunogen: Blocking, Flow cytometry, Inhibition). View Reference
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Gravina A, Tediashvili G, Rajalingam R, et al. Protection of cell therapeutics from antibody-mediated killing by CD64 overexpression.. Nat Biotechnol. 2023; 41(5):717-727. (Clone-specific: Cell separation, Flow cytometry). View Reference
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Indik ZK, Hunter S, Huang MM, et al. The high affinity Fc gamma receptor (CD64) induces phagocytosis in the absence of its cytoplasmic domain: the gamma subunit of Fc gamma RIIIA imparts phagocytic function to Fc gamma RI. Exp Hematol. 1994; 22(7):599-606. (Biology). View Reference
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Sun W, O'Shea JJ, Guyre PM. CD64 Workshop Panel Report. In: 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.; 1997:988-990.
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van Vugt MJ, Heijnen AF, Capel PJ, et al. FcR gamma-chain is essential for both surface expression and function of human Fc gamma RI (CD64) in vivo. Blood. 1996; 87(9):3593-3599. (Biology). View Reference
571787 Rev. 1
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.