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Multiparameter flow cytometric analysis using BD OptiBuild™ RB780 Mouse Anti-Human CD64 antibody (Cat. No. 755954) on human peripheral blood. Flow cytometry was performed using a BD LSRFortessa™ X-20 Flow Cytometer System.
BD OptiBuild™ RB780 Mouse Anti-Human CD64
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
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
- 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.
- Researchers should determine the optimal concentration of this reagent for their individual applications.
- The production process underwent stringent testing and validation to assure that it generates a high-quality conjugate with consistent performance and specific binding activity. However, verification testing has not been performed on all conjugate lots.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- An isotype control should be used at the same concentration as the antibody of interest.
- 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.
- Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
- 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.
Companion Products
The MD22 monoclonal antibody specifically recognizes CD64 which is also known as Fc-gamma receptor I (FcgR1) or FcγRI. CD64 is a ~72 kDa type I transmembrane glycoprotein that is encoded by FCGR1A (Fc fragment of IgG receptor Ia). CD64 functions as a high affinity receptor for human IgG. The CD64 antigen is expressed on monocytes, macrophages, at low levels on polymorphonuclear neutrophils (PMNs), and on a subpopulation of circulating dendritic cells. The CD64 antigen is an early granulomonocytic lineage marker on CD34+ hematopoietic progenitors. Expression of the CD64 antigen increases transiently in cases of sepsis. The CD64 antigen functions in both innate and adaptive immune responses, and mediates endocytosis, phagocytosis, antigen presentation, antibody-dependent cellular cytotoxicity, cytokine release, and superoxide generation. The CD64 antigen associates with the signal-transducing γ-chain homodimer of Fc receptors to form the functional high affinity FcγRI complex. Ligation of the CD64 antigen leads to the activation of the protein tyrosine kinases, hck and lyn.
Development References (10)
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Ernst LK, Duchemin AM, Anderson CL. Association of the high-affinity receptor for IgG (Fc gamma RI) with the gamma subunit of the IgE receptor.. Proc Natl Acad Sci USA. 1993; 90(13):6023-7. (Biology). View Reference
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Fanger NA, Wardwell K, Shen L, Tedder TF, Guyre PM. Type I (CD64) and type II (CD32) Fc gamma receptor-mediated phagocytosis by human blood dendritic cells.. J Immunol. 1996; 157(2):541-8. (Biology). View Reference
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Fischer G, Schneider EM, L Moldawer LL, et al. CD64 surface expression on neutrophils is transiently upregulated in patients with septic shock.. Intensive Care Med. 2001; 27(12):1848-52. (Biology). View Reference
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Olweus J, Terstappen LW, Thompson PA, Lund-Johansen F. Expression and function of receptors for stem cell factor and erythropoietin during lineage commitment of human hematopoietic progenitor cells. Blood. 1996; 88:1594-1607. (Biology). View Reference
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Qureshi SS, Lewis SM, Gant VA, Treacher D, Davis BH, Brown KA. Increased distribution and expression of CD64 on blood polymorphonuclear cells from patients with the systemic inflammatory response syndrome (SIRS).. Clin Exp Immunol. 2001; 125(2):258-65. (Biology). View Reference
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Scholl PR, Geha RS. Physical association between the high-affinity IgG receptor (Fc gamma RI) and the gamma subunit of the high-affinity IgE receptor (Fc epsilon RI gamma).. Proc Natl Acad Sci USA. 1993; 90(19):8847-50. (Biology). View Reference
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Wang AV, Scholl PR, Geha RS. Physical and functional association of the high affinity immunoglobulin G receptor (Fc gamma RI) with the kinases Hck and Lyn.. J Exp Med. 1994; 180(3):1165-70. (Biology). View Reference
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Zola H. Leukocyte and stromal cell molecules : the CD markers. Hoboken, N.J.: Wiley-Liss; 2007.
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van de Winkel JG, Anderson CL. Biology of human immunoglobulin G Fc receptors.. J Leukoc Biol. 1991; 49(5):511-24. (Biology). View Reference
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van de Winkel JG, Capel PJ. Human IgG Fc receptor heterogeneity: molecular aspects and clinical implications.. Immunol Today. 1993; 14(5):215-21. (Biology). View Reference
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.