Cd64 x54 5 7.1

Manufactured by BD

The CD64 (X54-5/7.1) is a laboratory equipment product. It is used for the detection and analysis of the CD64 cell surface antigen, also known as the high-affinity Fc gamma receptor I (FcγRI). The CD64 antigen is expressed on the surface of certain immune cells, such as monocytes and macrophages, and plays a role in the immune response.

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Lab products found in correlation

Siglecf e50 2440, by BD (2 mentions) Foxp3 fix perm buffer set, by BioLegend (1 mentions) Fc block rat anti mouse cd16 32, by Thermo Fisher Scientific (1 mentions) Anti foxp3 150d, by BioLegend (1 mentions) Gata 3 twaj, by Thermo Fisher Scientific (1 mentions) Facscanto 2 flow cytometer, by BD (1 mentions) Collagenase, by Merck Group (1 mentions) F4 80 bm8, by BioLegend (1 mentions) Cd45 30 f11, by Thermo Fisher Scientific (1 mentions) Cd206 c068c2, by BioLegend (1 mentions) Facsaria 2, by BD (1 mentions) Cd86 gl1, by Thermo Fisher Scientific (1 mentions) Cd103 2e7, by Thermo Fisher Scientific (1 mentions) Lsrfortessa, by BD (1 mentions) Alexa flour 488 conjugated goat anti rabbit igg, by Thermo Fisher Scientific (1 mentions) Foxp3 fix perm buffer, by BioLegend (1 mentions) Facscanto 2, by Tree Star (1 mentions) Cd31 390, by Thermo Fisher Scientific (1 mentions) Isotype control antibody, by BD (1 mentions)

Spelling variants of this product

CD64 PE (X54-5/7.1 monoclonal, (2 citations)

6 protocols using cd64 x54 5 7.1

Adipose Tissue Immune Cell Profiling

Cited 3 times

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Adipose tissue SVF isolation and flow cytometry were performed as previously described (46 (link), 47 (link)). Briefly, SVF was incubated in Fc Block (rat anti-mouse CD16/32; eBioscience) before staining with anti-CD45 (30-F11), CD3e (145-2C11), CD4 (GK1.5), CD8a (53-6.7), CD11c (N418), and CD64 (X54-5/7.1) (BD Pharmingen). For analysis of T_regs, the samples were fixed after surface staining, permeabilized with a FOXP3 Fix/Perm Buffer Set (421403; BioLegend), and stained with anti-FoxP3 (150D; BioLegend) antibody according to the manufacturer’s protocol. For ILC2 cells, SVF was stained with anti-CD45 (30-F11), B220 (RA3-6B2), CD11b (M1/70), CD11c (N418), Gr-1 (RB6-8C5), CD3e (145-2C11), Thy1.2 (53-2.1) (BioLegend), ST2 (RMST2-2), and Gata3 (TWAJ) (eBioscience), as previously described (11 (link)). Samples were analyzed using a BD LSR cell analyzer at the Vision Research Core Facility at the University of Michigan Medical School. Data were analyzed using the FACSDiva v6.2 software (BD Biosciences) and Flowjo (Flowjo.com). Flow cytometry gating strategies for T_regs, B cells, macrophages, neutrophils, CD8⁺ T cells, and ILC2s are shown in fig. S5.

Zhao X.Y., Zhou L., Chen Z., Ji Y., Peng X., Qi L., Li S, & Lin J.D. (2020). The obesity-induced adipokine sST2 exacerbates adipose Treg and ILC2 depletion and promotes insulin resistance. Science Advances, 6(20), eaay6191.

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Flow Cytometric Analysis of Liver Macrophages

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Livers were perfused with 1X PBS, mashed and digested in DMEM containing 250 μg/ml Collagenase (Sigma) at 37°C for 30 min. The digested livers were then mashed and filtered through a 100 μm metal strainer and digestion was repeated for 15 min. Total liver contents were strained and washed with DMEM. The pellet was lysed with 1X lysis buffer (BD PharmLyse), quenched, and washed. The resulting cell suspension was used in flow cytometry. Surface staining was performed using the following mAb against mouse antigens: CD45 (30-F11, eBioscience), CD301(BioRad), CD206 (C068C2, Biolegend), F4/80 (BM8, Biolegend), mouse Mer biotinylated (R&D), CD64(X54-5/7.1, BD). Samples were acquired using FACSCanto II flow cytometer (BD) and analyzed using Flowjo X 10.0.7r2 (FlowJo LLC, Inc.,).

Cortes-Selva D., Elvington A.F., Ready A., Rajwa B., Pearce E.J., Randolph G.J, & Fairfax K.C. (2018). Schistosoma mansoni Infection-Induced Transcriptional Changes in Hepatic Macrophage Metabolism Correlate With an Athero-Protective Phenotype. Frontiers in Immunology, 9, 2580.

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Multiparameter FACS Analysis of Immune Cell Subsets

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Floating cells and attached cells were separately prepared for FACS analysis. Cells were then incubated for 20 min with antibodies diluted at the optimal concentrations in FACS buffer (PBS, 5% FBS, 5 mM EDTA, and 1% NaN₃). LSRII (BD) was used for multiparameter analysis of stained cell suspensions, followed by analysis with FlowJo software (Tree Star) as described previously (Seok et al., 2013 (link)). Monoclonal antibodies to mouse F4/80 (BM8), CD11b (M1/70), IA/IE (M5/114.15.2), CD8α (53-6.7), CD115 (AFS98), FcεRI (MAR-1), Siglec F (E50-2440, BD Biosciences), Ly6C (HK1.4), MerTK (clone 125518, R&D systems), CD64 (X54-5/7.1, BD Biosciences), CD11c (HL3), CD80 (16-10A1, BD Biosciences), CD86 (GL1), and CD103 (2E7), were all from eBioscience, unless indicated. MHCII^highF4/80^low and MHCII^lowF4/80^high populations were sorted with a FACSAria II (BD). Sorting purity was always checked and confirmed that was up to 95~98%.

Na Y.R., Jung D., Gu G.J, & Seok S.H. (2016). GM-CSF Grown Bone Marrow Derived Cells Are Composed of Phenotypically Different Dendritic Cells and Macrophages. Molecules and Cells, 39(10), 734-741.

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Multiparametric Flow Cytometry Analysis

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Floating cells were prepared for FACS analysis. Cells were incubated for 20 min with antibodies diluted at the optimal concentrations in FACS buffer (PBS, 5% FBS, 5 mM EDTA, and 1% NaN₃). LSRFortessa (BD Biosciences) was used for multiparameter analysis of stained cell suspensions followed by analysis with FlowJo software (Tree Star Inc.). Monoclonal antibodies to mouse CD45 (30-F11), Ly6G (1A8-Ly6g), Ly6C (HK1.4), F4/80 (BM8), CD11b (M1/70), CD3 (145-2C11), CD8α (53-6.7), and CD64 (X54-5/7.1, BD Biosciences) were all from eBioscience unless otherwise indicated. Phospho-PDHE1α (NB110-93479; Novus Biologicals) staining was performed after cell fixation and permeabilization followed by secondary Alexa Flour 488-conjugated goat anti-rabbit IgG (Invitrogen) staining.

Na Y.R., Jung D., Song J., Park J.W., Hong J.J, & Seok S.H. (2020). Pyruvate dehydrogenase kinase is a negative regulator of interleukin-10 production in macrophages. Journal of Molecular Cell Biology, 12(7), 543-555.

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Adipose Tissue Immune Cell Analysis

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Adipose tissue was digested in RPMI with 0.5% BSA and 1 mg/ml type II collagenase on a rocking platform for 25 min at 37 °C. The SVF was separated from adipocytes by centrifugation. The following antibodies were used for flow cytometry following Fcblock: CD45 (30-F11), CD3e (145-2C11), CD4 (GK1.5), CD8a (53-6.7), CD11c (N418), Sca-1 (Ly-6A/E) (D7), CD31 (390) (eBioscience), PDGFRα (RM0004-3G28) (Abcam), and CD64 (X54-5/7.1) (BD Pharmingen). For intracellular collagen staining, cells were incubated in Fc Block then stained with surface antibodies for 45 min at 4°C. Intracellular stains were performed using FOXP3 Fix/Perm buffer (BioLegend, San Diego, CA) followed by 15 min block with 0.5% goat serum, a 1hr incubation with 0.5μg of rabbit Collagen Type I antibody (Rockland, Limerick, PA), three washes, and 30 min incubation with 0.2μg of goat anti-rabbit AlexaFluor 647 antibody (Thermo Fisher Scientific-Life Technologies, Carlsbad, CA). Analysis was performed using a BD Biosciences FACSCanto II and FlowJo v.10 (Treestar).

Zamarron B.F., Porsche C.E., Luan D., Lucas H.R., Mergian T.A., Martinez-Santibanez G., Cho K.W., DelProposto J.L., Geletka L.M., Muir L.A., Singer K, & Lumeng C.N. (2020). Weight Regain in Formerly Obese Mice Hastens the Development of Hepatic Steatosis Due To Impaired Adipose Tissue Function. Obesity (Silver Spring, Md.), 28(6), 1086-1097.

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Comprehensive Immune Cell Analysis

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Graft-infiltrating cells were assessed using CD45.2 (104), CD45.1 (A20), Gr1 (RB6-8C5), Ly6G (1A8), CD11c (N418), CD11b (M1/70), I-A^b (25-9-17), Siglec F (E50-2440), CD64 (X54-5/7.1) and isotype control antibodies (BD Biosciences, San Jose, CA BioLegend, San Diego, CA and eBiosciences, San Diego, CA).

Spahn J.H., Li W., Bribriesco A.C., Liu J., Shen H., Ibricevic A., Pan J., Zinselmeyer B.H., Brody S.L., Goldstein D.R., Krupnick A.S., Gelman A.E., Miller M.J, & Kreisel D. (2015). DAP12 expression in lung macrophages mediates ischemia reperfusion injury by promoting neutrophil extravasation. Journal of immunology (Baltimore, Md. : 1950), 194(8), 4039-4048.

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