Cells were blocked with anti-CD32/16 (clone 2.4G2, produced in-house) and stained with surface antibodies as indicated; CD8-PE (53-6.7), CD62L-APC (MEL-14), IFNγ-PE Cy7 (XMG1.2), CD11b APC CY7 (M1-70), and GR-1 FITC (RB6-8C5) from BD Pharmingen; B220-A647 (RA3-6B2), CD3-PE Cy7 (145-2C11), CD8-A700 (53-6.7), CD11c-eFluor450 (N418), CD44-PE Cy7 (IM7), CD45.1-PE (A20), Vα2-APC (B20.1) from eBioscience, CD11c-PE Cy7 (N418) from BioLegend. A viability dye, live/dead fixable blue (Invitrogen), was included before fixing cells with formalin (Sigma-Aldrich). Samples were collected on an LSRII SORP (BD, San Jose, CA, USA). Data were analyzed using FlowJo version 9.6 (Tree Star, Ashland, OR, USA).
Gr 1 fitc rb6 8c5
Manufactured by BD
Sourced in United States
The GR-1 FITC (RB6-8C5) is a fluorescently-labeled antibody that binds to the Gr-1 antigen, also known as Ly-6G and Ly-6C. It is commonly used in flow cytometry applications to identify and quantify granulocytes, including neutrophils, in cell samples.
Automatically generated - may contain errors
Lab products found in correlation
Cd11c pe cy7 n418, by BioLegend (1 mentions)
Lsrii sorp, by BD (1 mentions)
Live dead fixable blue, by Thermo Fisher Scientific (1 mentions)
Cd11b apc cy7 m1 70, by BD (1 mentions)
Formalin, by Merck Group (1 mentions)
Cd11c apc hl3, by BD (1 mentions)
Cd4 fitc gk1, by Thermo Fisher Scientific (1 mentions)
Cd11b apc m1 70, by BD (1 mentions)
Cd80 fitc 16 10a1, by Thermo Fisher Scientific (1 mentions)
Cd11b apc, by Thermo Fisher Scientific (1 mentions)
Cd8a apc, by Thermo Fisher Scientific (1 mentions)
Mhcii fitc, by Thermo Fisher Scientific (1 mentions)
Gr 1 fitc, by Thermo Fisher Scientific (1 mentions)
Cd11c apc, by Thermo Fisher Scientific (1 mentions)
Cd3 apc 145 2c11, by Thermo Fisher Scientific (1 mentions)
Dnase 1, by Roche (1 mentions)
Cd11b apc m1 70, by Thermo Fisher Scientific (1 mentions)
Ly6c pe hk1.4, by Thermo Fisher Scientific (1 mentions)
Collagenase type 1, by Worthington (1 mentions)
Cell strainer, by Corning (1 mentions)
4 protocols using gr 1 fitc rb6 8c5
1
Comprehensive Immune Cell Profiling
2
Multi-Marker Immune Cell Profiling
3
Isolation and Analysis of Tumor-Derived Myeloid Cells
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Primary tumors were isolated from mice, cut into pieces, and digested in RPMI1640 containing 200 U·mL−1 of collagenase, type I (Worthington, Lakewood, NJ, USA) and 10 μg·mL−1 of DNaseI (Roche Diagnostics) for 60 min at 37 °C on a shaking platform. The samples were then washed and filtered through a cell strainer (100‐μm nylon; Corning, Corning, NY, USA). Red blood cells were lysed in Red Blood Cell Lysis Buffer (Roche Diagnostics). The collected cells were incubated with FcR blocking reagent, mouse (Miltenyi Biotec, Bergisch Gladbach, Germany), for 15 min on ice. For flow cytometry, the cells were stained with the following antibodies for 30 min on ice in the dark: CD11b‐APC (M1/70) and Ly6C‐PE (HK1.4) from eBioscience (San Diego, CA, USA), Ly6G‐PE/Cy7 (1A8) and CD45‐Alexa Fluor 700 (30‐F11) from BioLegend (San Diego, CA, USA), and Gr‐1‐FITC (RB6‐8C5) from BD Biosciences (San Jose, CA, USA). The cells were analyzed by Gallios Flow Cytometer (Beckman Coulter, Fullerton, CA, USA) and were further analyzed using flowjo software (TreeStar software, Ashland, OR, USA).
4
VSV-Mediated Cytokine Induction in EMT6 Cells
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EMT6 cells were cotreated with 0.1 MOI of VSVΔ51-GFP and 5 μM LCL161 for 20 hr. Cells were trypsinized, permeabilized with the CytoFix/CytoPerm kit (BD Biosciences) and stained with APC-TNFα (MP6-XT22, BD Biosciences). Cells were analyzed on a Cyan ADP 9 flow cytometer (Beckman Coulter) and data was analyzed with FlowJo (Tree Star).
Splenocytes were enriched for CD11b using the EasySep CD11b positive selection kit (StemCell Technologies). CD49+ cells were enriched using the EasySep CD49b positive selection kit (StemCell Technologies) from the CD11b− fraction. CD11b+ cells were stained with F4/80-PE-Cy5 (BM8, eBioscience) and Gr1-FITC (RB6-8C5, BD Biosciences) and further sorted with MoFlo Astrios (Beckman Coulter). Flow cytometry data was analyzed using Kaluza (Beckman Coulter). Isolated cells were infected with VSVΔ51 for 24 hr and clarified cell culture supernatants was applied to EMT6 cells for 24 hr in the presence of 5 μM LCL161. An n = 3 of biological replicates was used to determine statistical measures (mean, standard deviation).
Splenocytes were enriched for CD11b using the EasySep CD11b positive selection kit (StemCell Technologies). CD49+ cells were enriched using the EasySep CD49b positive selection kit (StemCell Technologies) from the CD11b− fraction. CD11b+ cells were stained with F4/80-PE-Cy5 (BM8, eBioscience) and Gr1-FITC (RB6-8C5, BD Biosciences) and further sorted with MoFlo Astrios (Beckman Coulter). Flow cytometry data was analyzed using Kaluza (Beckman Coulter). Isolated cells were infected with VSVΔ51 for 24 hr and clarified cell culture supernatants was applied to EMT6 cells for 24 hr in the presence of 5 μM LCL161. An n = 3 of biological replicates was used to determine statistical measures (mean, standard deviation).
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