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Buv563

Manufactured by BD

BUV563 is a fluorochrome-conjugated antibody reagent designed for flow cytometry applications. It is used to detect and analyze specific cell surface markers expressed on cells.

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4 protocols using buv563

1

Profiling Immune Cell Populations in Inguinal Lymph Nodes

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Draining inguinal lymph nodes from immunized mice were collected and treated as previously described (47 (link), 48 (link)). Briefly, the tissues were treated with collagenase type IV (Worthington) at a concentration of 1 mg/mL for 20 min at a temperature of 37 °C, and then passed through a 100-μm strainer to obtain a single-cell suspension. The resulting single-cell samples were stained with a range of markers including Zombie UV (BUV496, BioLegend 423107), anti-Ly6C (BV780, BioLegend 128041), anti-Ly6G (APC-Cy7, BioLegend 127624), anti-CD19 (BUV395, BD 563557), anti-CD3 (BB700, BD742175), anti-MHCII (AF700, eBioscience 56-5321-82), anti-CD11b (BV650, BioLegend 101239), anti-CD11c (BV421, BioLegend 117330), anti-CD86 (A647, BioLegend 105020), anti-Siglec-F (PE-CF594, BD 562757), anti-CD45 (BV610, BioLegend 103140), anti-CD169 (PE-Cy7, BioLegend 142412), anti-PDCA-1 (BUV563, BD 749275), anti-CD8a (BUV805, BD 612898), anti-CD103 (PE, eBioscience 12-1031-82), anti-NK1.1 (BV510, BioLegend 108738), and anti-F4/80 (BUV737, BD 749283). The cells were analyzed using the BD FACSymphony analyzer located at the Stanford Shared Fluorescence-activated cell sorting (FACS) Facility.
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2

Comprehensive Immune Cell Profiling of Vaccinated Mice

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Draining iliac lymph nodes from BNT162b2 immunized mice, draining inguinal lymph nodes from YF-17D immunized mice, along with whole lung or spleen, were harvested and digested with 1 mg/mL collagenase type IV (Worthington) for 20 min at 37 °C, followed by smashing with a 100 μm strainer to make a single-cell suspension. Red blood cells from the lung and spleen were lysed before staining. Single-cell samples were then stained with Zombie UV™ (1:300, BUV496; BioLegend #423107), anti-Ly6C (1:500, BV780; BioLegend #128041), anti-Ly6G (1:400, APC-Cy7; BioLegend #127624), anti-CD19 (1:100, BB700; BD #566411), anti-CD3 (1:100, BB700; BD #742175), anti-MHCII (1:400, AF700; eBioscience #56-5321-82), anti-CD11b (1:300, BV650; BioLegend #101239), anti-CD11c (1:400, BV421; BioLegend #117330), anti-CD86 (1:300, A647; BioLegend #105020), anti-Siglec-F (1:400, PE-CF594; BD #562757), anti-CD45 (1:200, BV610; BioLegend #103140), anti-CD169 (1:200, PE-Cy7; BioLegend #142412), anti-PDCA-1 (1:200, BUV563; BD #749275), anti-CD8a (1:200, BUV805; BD #612898), anti-CD103 (1:100, PE; eBioscience #12-1031-82), anti-NK1.1 (1:200, BV510; BioLegend #108738), and anti-F4/80 (1:100, BUV737; BD #749283). Data was collected on a BD FACSymphony analyzer with BD FACSDiva (v8.0.1).
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3

Comprehensive Immune Profiling of Iliac Lymph Nodes

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Iliac lymph nodes were harvested at day 1 post-vaccination. LNs were treated with 5 mg/ml collagenase type IV (Worthington) for 20 min at 37 °C and smashed against a 100μm strainer to make a single-cell suspension. Cells were then stained with Zombie UV™ (BUV496; Biolegend # 423108, 1:250 dilution), anti-CD64 (A488; Biolegend # 139316, 1:100 dilution), anti-Ly6C (BV780; Biolegend # 128041, 1:500 dilution), anti-Ly6G (APC-Cy7; Biolegend# 127624, 1:400 dilution), anti-CD19 (BB700; BD # 566411, 1:200 dilution), anti-CD3 (BB700; BD #742175, 1:200 dilution), anti-MHCII (AF700; eBioscience #56-5321-82, 1:400 dilution), anti-CD11b (BV650; Biolegend #101239, 1:300 dilution), anti-CD11c (BV421; Biolegend #117330, 1:400 dilution), anti-CD86 (A647; Biolegend #105020, 1:300 dilution), anti-Siglec-F (PE-CF594; BD #562757, 1:400 dilution), anti-CD24 (BUV395; BD #744471, 1:200 dilution), anti-CD45 (BV610; Biolegend #103140, 1:300 dilution), anti-CD169 (PE-Cy7; Biolegend #142412, 1:200 dilution), anti-PDCA-1 (BUV563; BD #749275, 1:200 dilution), anti-CD8a (BUV805; BD #612898, 1:200 dilution), anti-CD103 (PE; eBioscience #12-1031-82, 1:100 dilution), anti-NK1.1 (BV510; Biolegend #108738, 1:200 dilution), anti-F4/80 (BUV737; BD #749283, 1:100 dilution). Cells were analyzed with the BD FACSymphony analyzer at Stanford HIMC core.
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4

Comprehensive Immune Cell Profiling of Vaccinated Mice

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Draining iliac lymph nodes from BNT162b2 immunized mice, draining inguinal lymph nodes from YF-17D immunized mice, along with whole lung or spleen, were harvested and digested with 1 mg/mL collagenase type IV (Worthington) for 20 min at 37 °C, followed by smashing with a 100 μm strainer to make a single-cell suspension. Red blood cells from the lung and spleen were lysed before staining. Single-cell samples were then stained with Zombie UV™ (1:300, BUV496; BioLegend #423107), anti-Ly6C (1:500, BV780; BioLegend #128041), anti-Ly6G (1:400, APC-Cy7; BioLegend #127624), anti-CD19 (1:100, BB700; BD #566411), anti-CD3 (1:100, BB700; BD #742175), anti-MHCII (1:400, AF700; eBioscience #56-5321-82), anti-CD11b (1:300, BV650; BioLegend #101239), anti-CD11c (1:400, BV421; BioLegend #117330), anti-CD86 (1:300, A647; BioLegend #105020), anti-Siglec-F (1:400, PE-CF594; BD #562757), anti-CD45 (1:200, BV610; BioLegend #103140), anti-CD169 (1:200, PE-Cy7; BioLegend #142412), anti-PDCA-1 (1:200, BUV563; BD #749275), anti-CD8a (1:200, BUV805; BD #612898), anti-CD103 (1:100, PE; eBioscience #12-1031-82), anti-NK1.1 (1:200, BV510; BioLegend #108738), and anti-F4/80 (1:100, BUV737; BD #749283). Data was collected on a BD FACSymphony analyzer with BD FACSDiva (v8.0.1).
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