Cells were suspended in 3% FBS 2mM EDTA in PBS and staining was performed in the presence of 2% NRS, 2% Fc block (BD), and fixable viability dye eFluor 450 (eBioscience). Cells were stained with the following antibodies from BD: CD3-APC (145-2C11), CD19- APC (1D3), CD11c-V450 or PE-Cy7 (HL3), NK1.1-APC (PK136), Ly6G-V450 (1A8), Ly6C-PerCP-Cy5.5 (AL-21), CD11b-PE (M1/70), B220-APC (RA3-6B2), CD45-APC (30-F11), SiglecF-BV421 (E50-2440), CD8-PerCP-Cy5.5 (53-6.7). And the following antibodies from eBioscience: CD103-APC (2E7), EpCAM-PE-Cy7 (G8.8), CD31-PerCP-eFluor 710 (390), MHC Class II (I-A/I-E) eFluor 450 (M5/114.15.2), FasL-APC (MFL3), CD4-APC (GK1.5), and from R&D: CCR2-APC (475301). Alexa Fluor 647 protein labeling kit (Thermo) was used to label the influenza M2 (E10) (Bourmakina and García-Sastre, 2005 (link)) antibody. Cells were fixed with 2% formaldehyde after staining and analyzed on an LSRII after gating for FSC/SSC, singlets, and live cells. Cells were quantitated by flow cytometry with AccuCount Particles (Spherotech). Sorting was performed similarly on FACSAria.
Efluor 450
Manufactured by R&D Systems
EFluor 450 is a fluorescent dye that can be used for cell staining and analysis. It has an excitation maximum of 450 nm and an emission maximum of 550 nm, allowing detection in the blue-green region of the visible spectrum.
Automatically generated - may contain errors
Lab products found in correlation
Fc block, by BD (2 mentions)
Efluor 450, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 647 protein labeling kit, by Thermo Fisher Scientific (2 mentions)
Cd8 percp cy5, by BD (2 mentions)
Ly6c percp cy5, by BD (2 mentions)
Cd3 apc 145 2c11, by BD (2 mentions)
Cd4 apc, by R&D Systems (2 mentions)
Ccr2 apc, by R&D Systems (2 mentions)
Cd11c v450 or pe cy7 hl3, by BD (2 mentions)
Siglecf bv421 e50 2440, by BD (2 mentions)
Cd31 percp efluor 710 390, by R&D Systems (2 mentions)
Fasl apc, by R&D Systems (2 mentions)
Accucount particles, by Spherotech (2 mentions)
2 protocols using efluor 450
1
Multicolor Flow Cytometry Analysis
2
Multicolor Flow Cytometry Analysis
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Cells were suspended in 3% FBS 2mM EDTA in PBS and staining was performed in the presence of 2% NRS, 2% Fc block (BD), and fixable viability dye eFluor 450 (eBioscience). Cells were stained with the following antibodies from BD: CD3-APC (145-2C11), CD19- APC (1D3), CD11c-V450 or PE-Cy7 (HL3), NK1.1-APC (PK136), Ly6G-V450 (1A8), Ly6C-PerCP-Cy5.5 (AL-21), CD11b-PE (M1/70), B220-APC (RA3-6B2), CD45-APC (30-F11), SiglecF-BV421 (E50-2440), CD8-PerCP-Cy5.5 (53-6.7). And the following antibodies from eBioscience: CD103-APC (2E7), EpCAM-PE-Cy7 (G8.8), CD31-PerCP-eFluor 710 (390), MHC Class II (I-A/I-E) eFluor 450 (M5/114.15.2), FasL-APC (MFL3), CD4-APC (GK1.5), and from R&D: CCR2-APC (475301). Alexa Fluor 647 protein labeling kit (Thermo) was used to label the influenza M2 (E10) (Bourmakina and García-Sastre, 2005 (link)) antibody. Cells were fixed with 2% formaldehyde after staining and analyzed on an LSRII after gating for FSC/SSC, singlets, and live cells. Cells were quantitated by flow cytometry with AccuCount Particles (Spherotech). Sorting was performed similarly on FACSAria.
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