Bv605 ly6 c

Manufactured by BioLegend

BV605-Ly6-C is a fluorochrome-conjugated antibody used for the detection and analysis of Ly6-C, a cell surface antigen, in flow cytometry applications. The BV605 fluorochrome provides a bright signal for sensitive detection.

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3 protocols using bv605 ly6 c

Multiparametric Flow Cytometry Analysis of Murine Splenocytes

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An LSR Fortessa (BD Biosciences, Franklin Lakes, NJ) was used for multiparametric flow cytometry analysis. A Live/Dead Fixable Blue Dead Cell Stain Kit (Molecular Probes; Thermo Fisher Scientific, Grand Island, NY) was used to exclude dead cells. The following murine monoclonal antibodies (mAbs) were used to stain Balb/c splenocytes to characterize immune cell subsets: FITC-CD8 (Clone 53-6.7; BD Biosciences), PE-CD49b (Clone DX5; BioLegend, San Diego, CA), PE-PD-L1 (Clone 10F.9G2; BioLegend), PerCP-Cy5.5-B7-1 (Clone 16-10A1; BioLegend), PerCP-Cy5.5-FOXP3 (Clone FJK-16s; eBioscience, San Diego, CA), PE-Cy7-CD122 (Clone Tm-b1; eBioscience), APC-NKG2D (Clone CX5; BioLegend), APC-Cy7-CD3 (Clone 17A2; BioLegend), Pacific Blue-CD44 (Clone IM7; BioLegend), AF700-CD4 (Clone GK1.5; eBioscience), BV605-CD19 (Clone 6D5; BioLegend), BV605-PD-1 (Clone 29F.1A12; BioLegend), BV510-NKp46 (Clone 29A1.4; BioLegend), Pacific Blue-CD27 (Clone LG.3A10; BioLegend), FITC-CD11b (Clone M1/70; BD Biosciences), AF700-CD11b (Clone M1/70; BioLegend), APC-Cy7-CD11c (Clone N418; BioLegend), BV510-CD3 (Clone 17A2; BioLegend), Pacific Blue-Ly6-G (Clone 1A8; BioLegend), BV605-Ly6-C (Clone HK1.4; BioLegend).

Kim P.S., Kwilas A.R., Xu W., Alter S., Jeng E.K., Wong H.C., Schlom J, & Hodge J.W. (2016). IL-15 superagonist/IL-15RαSushi-Fc fusion complex (IL-15SA/IL-15RαSu-Fc; ALT-803) markedly enhances specific subpopulations of NK and memory CD8+ T cells, and mediates potent anti-tumor activity against murine breast and colon carcinomas. Oncotarget, 7(13), 16130-16145.

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Comprehensive Immune Profiling of PDAC Tumors

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PDAC cells were detached using Cell Dissociation Buffer (ThermoFisher, #13151014), then stained with PE-conjugated HLA-A, B, C antibodies (1:200, Biolegend#311405) for 1 hour. Viable cells were stained with DAPI. For immune profiling of tumors, cells were isolated from the tumor using a 70 μm strainer. Isolated cells were stained with APC/Cy7-CD3(#10222), APC-CD45(103112), PE/Dazzle-594-CD8(#100762), PE-Cy5-CD4(#100514), PE-FoxP3(#126404), PerCP-Cy5.5-Cd11b (#101228), BV605-Ly6C (#128036), AF700-Ly6 (#127622) (all 1:200 dilution; Biolegend), and cell surface expression of these proteins were evaluated by flow cytometry. For autophagic flux determined by flow cytometry, the ratio of GFP:mCherry was calculated by comparing the percentages of GFP+ and mCherry+ cells. Analysis of all flow cytometry experiments was performed with Flowjo V10.

McBrearty N., Cho C., Chen J., Zahedi F., Peck A.R., Radaelli E., Assenmacher C.A., Pavlak C., Devine A., Yu P., Lu Z., Zhang H., Li J., Pitarresi J.R., Astsaturov I., Cukierman E., Rustgi A.K., Stanger B.Z., Rui H, & Fuchs S.Y. (2023). Tumor suppressive and immune-stimulating roles of cholesterol 25-hydroxylase in pancreatic cancer cells. Molecular cancer research : MCR, 21(3), 228-239.

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Immunophenotyping Mouse Blood Cell Populations

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Blood was collected from mice via the retro-orbital sinus, and red blood cells were lysed before staining with the following fluorochrome-conjugated antibodies: BV650 CD45.1 (BioLegend clone A20), AlexaFluor700 CD45.2 (BioLegend clone 104), BUV496 B220 (BD Biosciences clone RA3-6B2), PerCP-Cy5.5 CD3e (BioLegend clone 145-2C11), APC-Cy7 CD11b (BioLegend clone M1/70), APC Ly6g (BioLegend clone 1A8), BV605 Ly6c (BioLegend clone HK1.4), BV421 Ter-119 (BioLegend clone TER-119), and PE-Cy7 F4/80 (Invitrogen BM8). Data were collected using a LSRII (BD Biosciences) and analyzed using FlowJo V10 (BD Biosciences). The following cell surface markers were used to isolate or phenotype the following cell types: hematopoietic stem cells (HSCs), Lin-Sca-1+ c-Kit+ Flt3-CD150+ CD48-; HSPCs, Lin-Sca-1+ c-Kit+; granulocyte/macrophage-primed multipotent progenitors (MPP G/M ), Lin-Sca-1+ c-Kit+ Flt3-CD150-CD48+; common myeloid progenitor cells (CMPs) Lin-Sca-1-c-Kit+ CD34+ FcγR-; and granulocyte-macrophage progenitors (GMPs), Lin-Sca-1-c-Kit+ CD34+ FcγR+. Data were collected using a BD FACSymphony A5 or cells were prospectively isolated using a FACSymphony S6 (BD Biosciences). All flow cytometry data were analyzed using FlowJo V10.

Schwartz L.S., Young K.A., Stearns T.M., Boyer N., Mujica K.D, & Trowbridge J.J. (2024). Transcriptional and functional consequences of Oncostatin M signaling on young Dnmt3a-mutant hematopoietic stem cells. Experimental hematology, 130.

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