Colon collected from control or DSS-treated mice was cut into small pieces then digested with 2.5 mg/mL Collagenase D and 30 ng/mL DNAse1 for 40 min at 37 °C then filtered through a 70 µm filter to generate single cell suspension for FACS analyses. Cells were then stained with Fixable Viability Dye eFluor 506 (eBioscience, 65-0866-14), blocked with anti-mouse CD16/32 (eBioscience, 14016185), followed by staining with antibody cocktails for preadipocytes or immune cells. The antibody cocktail for immune cells includes FITC -CD45 (BioLegend, 103107), PECy7-CD11b (BioLegend, 101216), FITC-Ly6G (eBioscience, 11593182), PE-F4/80 (eBioscience,12480182), APC-CD11C (BioLegend, 117310), AF700-MHCII (eBioscience, 56532182), and APC-eFluro-CD4 (eBioscience, 47-0042-80) PE -CD19 (BioLegend, 115507). All antibodies were used at a final dilution of 1 to 100. FACS analyses for surface expression of immune cell markers were performed by the BD FACSCanto RUO machine and analyzed by FlowJo V10 software.
Af700 mhcii
Manufactured by Thermo Fisher Scientific
The AF700-MHCII is a laboratory instrument designed for the detection and analysis of major histocompatibility complex class II (MHC-II) proteins. It utilizes fluorescent labeling and advanced optical detection technology to provide precise measurement of MHC-II expression levels in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Ly6g fitc, by Thermo Fisher Scientific (2 mentions)
Fixable viability dye efluor 506, by Thermo Fisher Scientific (2 mentions)
F4 80 pe, by Thermo Fisher Scientific (2 mentions)
Cd11b pe cy7, by BioLegend (2 mentions)
Cd11c apc, by BioLegend (2 mentions)
Anti mouse cd16 32, by Thermo Fisher Scientific (2 mentions)
Cd45 fitc, by BioLegend (1 mentions)
Cd19 pe, by BioLegend (1 mentions)
2 protocols using af700 mhcii
1
Isolation of Immune Cells from Murine Colon
2
Mouse Ear Immune Cell Profiling
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Mouse ears were collected following euthanasia in cold complete RPMI media on ice and cut into small pieces within 60 min, then digested with 2.5 mg/ml collagenase D, and 30 ng/ml DNase1 for 2 h at 37 °C and then filtered through a 30 μm filter to generate single-cell suspension for fluorescence-activated cell sorting analyses. Cells were then stained with Fixable Viability Dye eFluor 506 (eBioscience; 65-0866-14) blocked with antimouse CD16/32 (eBioscience; 14016185, 1:50 dilution), followed by staining with antibody cocktails for immune cells. The antibody cocktail for immune cells included PECy7-CD11b (BioLegend; 101216, 1:200 dilution), FITC-Ly6G (eBioscience; 11593182, 1:150 dilution), PE-F4/80 (eBioscience; 12480182, 1:200 dilution), APC-CD11C (BioLegend; 117310, 1:100 dilution), AF700-MHCII (eBioscience; 56532182, 1:100 dilution), and c-Kit (Biolegend; 10581, 1:100 dilution). Fluorescence-activated cell sorting analyses for surface expression of immune cell markers were performed by the Biorad ZE5 machine and analyzed by FlowJo V10 software (BD Biosciences). Dead cells were gated out from the analyses.
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