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Ly6c pe cy7 clone hk1

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Ly6C-PE Cy7 (clone HK1.4) is a fluorochrome-conjugated antibody used for the detection and analysis of Ly6C expression on cells. This antibody binds to the Ly6C surface antigen and can be used in flow cytometry applications.

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Lab products found in correlation

Il 1α pe clone alf 161, by BioLegend (4 mentions) Cytofix cytoperm, by BD (2 mentions) Ly6g pe, by BioLegend (2 mentions) Ly6g bv510, by BioLegend (2 mentions) Pe f4 80 clone bm8, by BioLegend (2 mentions) Wga 488, by Thermo Fisher Scientific (2 mentions) Cd63 apc clone nvg 2, by BioLegend (2 mentions) Clone mz3, by BioLegend (2 mentions) F4 80 fitc clone bm8, by BioLegend (2 mentions) Cd11b pe clone m1 70, by BioLegend (2 mentions) Cd9 apc, by BioLegend (2 mentions) Il 1β apc, by Thermo Fisher Scientific (2 mentions) Cd11c bv605, by BioLegend (2 mentions) F4 80 fitc, by BioLegend (2 mentions) Cytofix cytoperm kit, by BD (2 mentions) Ly6g bv510 clone 1a8, by BioLegend (2 mentions) Cd11b alexa fluor 700 clone m1 70, by Thermo Fisher Scientific (1 mentions) Fortessa flow cytometer, by BD (1 mentions) Ly6g pe clone 1a8, by BioLegend (1 mentions) Cd64 apc clone x54 5 7, by BioLegend (1 mentions)

Spelling variants of this product

Ly6C (HK1.4, (34 citations) Ly6C (clone HK1.4, (26 citations) Ly6C-PE-Cy7 (22 citations) Ly6C-PE (14 citations) Clone HK1.4 (5 citations) PE-Cy7-conjugated Ly6C (clone HK1.4, (2 citations)

6 protocols using ly6c pe cy7 clone hk1

1

Multiparametric Immune Cell Analysis

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Cells were stained using the following fluorophore conjugated anti-mouse antibodies: WGA-488 (Invitrogen), Ly6G-FitC (clone 1A8, BioLegend), Ly6G-BV510 (clone 1A8, BioLegend), Ly6G-PE (clone 1A8, BioLegend), CD11c-BV605 (clone N418, BioLegend), F4/80-FitC (clone BM8, BioLegend), F4/80-PE (clone BM8, BioLegend), CD11b-PE (clone M1/70, BioLegend), Ly6C-PE Cy7 (clone HK1.4, BioLegend), CD63-APC (clone NVG-2, BioLegend), CD9-APC (clone MZ3, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), and IL-1β-APC (clone NJTEN3, ThermoFisher Scientific). Antibodies were diluted in wash buffer (PBS with 1% BSA and 2 mM EDTA). Cells were stained for 20 minutes at 4°C, washed with wash buffer, fixed for 15 minutes in BD Biosciences cytofix/cytoperm, and permeabilization prior to intracellular staining. ACEA Novocyte was used for flow cytometry, and Novoexpress software was used for subsequent analysis. AMNIS ImageStream was used for imaging flow cytometry and the AMNIS IDEAS software was used to calculate the colocalization coefficient.
Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.
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2

Flow Cytometric Analysis of Lung Immune Cells

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Lungs from mice subjected to CS or air were digested with collagenase as described (Stolberg et al, 2014 (link)) and single cell suspensions were labeled with antibodies for CD11c-V450 (clone N418; eBioscience), CD11b-AlexaFluor 700 (clone M1/70; eBioscience), MerTK-FITC (clone 2B10C42; BioLegend), CD64-APC (clone X54-5/7.1; BioLegend), Ly6C-PE/Cy7 (clone HK1.4; BioLegend), Ly6G-PE (clone 1A8; BioLegend), and CD103-PerCP/Cy5.5 (clone 2E7; BioLegend) at 4°C for 1 h. The population frequencies of resident AMs (CD11b−CD11c+ or MerTK+CD64+), neutrophils (CD11b+ Ly6G+), recruited monocytes (CD11c−CD11b+Ly6C+), and dendritic cells (CD11c+CD11b−CD103+) were analyzed on a BD Fortessa flow cytometer using BD FACSDiva software.
Penke L.R., Speth J.M., Draijer C., Zaslona Z., Chen J., Mancuso P., Freeman C.M., Curtis J.L., Goldstein D.R, & Peters-Golden M. (2020). PGE2 accounts for bidirectional changes in alveolar macrophage self-renewal with aging and smoking. Life Science Alliance, 3(11), e202000800.
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3

Quantification of Immune Cell Populations

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Total cell numbers were counted, and cells were stained using mouse Ly6G-BV510 (clone 1A8, BioLegend), Ly6C-PE-Cy7 (clone HK1.4, BioLegend), and F4/80-FitC (clone BM8, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), IL-1β-APC antibodies (clone NJTEN3, Thermo Fisher), and amine-reactive fixable viability dye e780 (Invitrogen, Thermofisher). Cell surface staining was performed at 4°C for 20 minutes. Cells were fixed for 15 minutes at 4°C and then permeabilized for intracellular stain (30 minutes) with Cytofix/Cytoperm kit (BD Biosciences). Flow cytometry and analysis was conducted on ACEA Novocyte instrument and Novoexpress software, respectively. The frequency of Ly6G+ neutrophils, F4/80+ macrophages, and F4/80- Ly6G- Ly6Chi monocytes were multiplied by total cell count to get cell numbers of each cell type.
Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.
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4

Multiparametric Immune Cell Analysis

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Cells were stained using the following fluorophore conjugated anti-mouse antibodies: WGA-488 (Invitrogen), Ly6G-FitC (clone 1A8, BioLegend), Ly6G-BV510 (clone 1A8, BioLegend), Ly6G-PE (clone 1A8, BioLegend), CD11c-BV605 (clone N418, BioLegend), F4/80-FitC (clone BM8, BioLegend), F4/80-PE (clone BM8, BioLegend), CD11b-PE (clone M1/70, BioLegend), Ly6C-PE Cy7 (clone HK1.4, BioLegend), CD63-APC (clone NVG-2, BioLegend), CD9-APC (clone MZ3, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), and IL-1β-APC (clone NJTEN3, ThermoFisher Scientific). Antibodies were diluted in wash buffer (PBS with 1% BSA and 2 mM EDTA). Cells were stained for 20 minutes at 4°C, washed with wash buffer, fixed for 15 minutes in BD Biosciences cytofix/cytoperm, and permeabilization prior to intracellular staining. ACEA Novocyte was used for flow cytometry, and Novoexpress software was used for subsequent analysis. AMNIS ImageStream was used for imaging flow cytometry and the AMNIS IDEAS software was used to calculate the colocalization coefficient.
Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.
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5

Quantification of Immune Cell Populations

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total cell numbers were counted, and cells were stained using mouse Ly6G-BV510 (clone 1A8, BioLegend), Ly6C-PE-Cy7 (clone HK1.4, BioLegend), and F4/80-FitC (clone BM8, BioLegend), IL-1α-PE (clone ALF-161, BioLegend), IL-1β-APC antibodies (clone NJTEN3, Thermo Fisher), and amine-reactive fixable viability dye e780 (Invitrogen, Thermofisher). Cell surface staining was performed at 4°C for 20 minutes. Cells were fixed for 15 minutes at 4°C and then permeabilized for intracellular stain (30 minutes) with Cytofix/Cytoperm kit (BD Biosciences). Flow cytometry and analysis was conducted on ACEA Novocyte instrument and Novoexpress software, respectively. The frequency of Ly6G+ neutrophils, F4/80+ macrophages, and F4/80- Ly6G- Ly6Chi monocytes were multiplied by total cell count to get cell numbers of each cell type.
Ratitong B., Marshall M, & Pearlman E. (2021). β-Glucan-stimulated neutrophil secretion of IL-1α is independent of GSDMD and mediated through extracellular vesicles. Cell reports, 35(7), 109139.
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6

Immune Cell Profiling in Mice

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Samples were Fc-blocked using anti-CD16/32 antibody (TruStainFcX, clone 93, BioLegend, 10 µg/ml, Catalog # 101320) before fluorescent antibody staining with Ly6G-Brilliant Violet 421 (clone 1A8, Biolegend, 0.5 µg/ml, Catalog # 127601), CD45.2-PerCP/Cy5.5 (clone104, Biolegend, 0.5 µg/ml, Catalog # 109827), Ly6C-PE/Cy7 (Clone HK 1.4, Biolegend, 2 µg/ml, Catalog # 128017), CD11b-APC/Cy7 (clone M1/70, Biolegend, 0.5 µg/ml, Catalog # 101225) and CD11c-APC (clone N418, Biolegend, 1 µg/ml, Catalog # 11730925). Analysis was performed with Fortessa or FACSAria (BD Biosciences) flow cytometers/sorters equipped with excitation lasers at 405 nm, 488 nm, 560 nm, and 633 nm. Data were analyzed by using the FlowJo 10.6.2 software (FlowJo, LLC).
Kleinholz C.L., Riek-Burchardt M., Seiß E.A., Amore J., Gintschel P., Philipsen L., Bousso P., Relja B., Schraven B., Handschuh J., Mohr J, & Müller A.J. (2021). Ly6G deficiency alters the dynamics of neutrophil recruitment and pathogen capture during Leishmania major skin infection. Scientific Reports, 11, 15071.
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