As described above, after Percoll separation, the cells were carefully collected and washed with cold FACS buffer and then stained for 30min at 4°C with anti-mouse-FITC-Gr1, anti-mouse-FITC-CD3e, anti-mouse-FITC-CD11b, anti-mouse-FITC-CD45R, anti-mouse-APC/Cy7-CD45, anti-mouse-PE-KLRG1 (1:500; 138408; BioLegend) and anti-mouse-APC-CD90 (1:500; 105312; BioLegend) antibodies. Lineage−CD45+KLRG1−CD90++ cells (ILC3) were sorted to ~97% purity using a FACS Aria II flow cytometer. ‘Lineage’ refers to the following antigens: Gr1, CD3e, CD11b and CD45R. A total of 10,000 cells per well were cultured in 96-well plates for 12h at 37°C in U-bottom plates (in RPMI-1640 medium). The reagent concentrations used for the treatment were as follows: 40ngml−1 mouse recombinant IL-23 (1887-ML; R&D Systems), 100μM GDCA (G9100; Sigma–Aldrich), 100μM TUDCA (T0266; Sigma–Aldrich), 10μM triamterene (PHR1722; Sigma–Aldrich) or the same dose of dimethyl sulfoxide (DMSO; D2650; Sigma–Aldrich). After 72h, the supernatants were collected for IL-22 detection, and the cells were collected for mRNA extraction.
Anti mouse fitc gr1
Manufactured by BioLegend
Sourced in United States
Anti-mouse-FITC-Gr1 is a fluorescently labeled antibody that binds to the Gr1 antigen expressed on the surface of mouse cells. The FITC fluorescent label allows for the detection and analysis of Gr1-positive cells using flow cytometry or other fluorescence-based techniques.
Automatically generated - may contain errors
Lab products found in correlation
Cck 8 kit, by Beyotime (2 mentions)
Tween 20, by Thermo Fisher Scientific (2 mentions)
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Bovine serum albumin (bsa), by Beyotime (2 mentions)
Paraformaldehyde (pfa), by Beyotime (2 mentions)
Electrophoresis solution, by Beyotime (2 mentions)
Transfer solution, by Beyotime (2 mentions)
Anti mouse percp cy5.5 ly6c, by BioLegend (2 mentions)
Tbs buffer, by Thermo Fisher Scientific (2 mentions)
Apc anti mouse ly6g, by BioLegend (2 mentions)
Fitc anti mouse cd4, by BioLegend (2 mentions)
Apc anti mouse cd25, by BioLegend (2 mentions)
Pe anti mouse cd11b, by BioLegend (2 mentions)
Dmem medium, by Thermo Fisher Scientific (2 mentions)
Rt pcr primers, by Sangon (2 mentions)
Tudca, by Merck Group (1 mentions)
D2650, by Merck Group (1 mentions)
Fitc anti mouse cd11b, by BioLegend (1 mentions)
Tumor dissociation kit, by Miltenyi Biotec (1 mentions)
3 protocols using anti mouse fitc gr1
1
Purification and Stimulation of ILC3s
2
Tumor and Spleen Cell Isolation Protocol
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The DMEM medium, PBS medium, and 10% fetal bovine serum were purchased from Gibco Life Technologies (Grand Island, NY, USA). The tumor dissociation kit and spleen dissociation kit were obtained from Miltenyi Biotec (San Diego, CA, USA). Anti-mouse PerCP/Cy5.5 CD3, anti-mouse FITC CD4, anti-mouse PE CD8, anti-mouse APC CD25, anti-mouse FITC Gr-1, anti-mouse PE CD11b, anti-mouse PerCP/Cy5.5 Ly6c, and anti-mouse APC Ly6g were all purchased from Biolegend (San Diego, CA, USA). 5%BSA, paraformaldehyde, electrophoresis solution, transfer solution, CCK-8 kit, RIPA were all provided by Beyotime Biotech (Beijing, China). The SDS-PAGs (Sodium Dodecyl Sulfate-Polyacrylamide Gels) were bought from Dakewe Biotech (Shenzhen, China). RT-PCR primers were designed and provided by Sangon Biotech (Shanghai, China). Trizol reagent, Tween-20, and 20 × TBS buffer were obtained from Thermo Scientific (Rockford, IL). Antibodies for Western blot were all purchased from Cell Signaling Technology (MA, USA). An ECL Kit was provided by Tanon Biotech (Shanghai, China).
3
Analyzing Gemcitabine's Apoptotic Effects
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The DMEM medium and 10% FBS were obtained from Gibco Life Technologies (Grand Island, NY, USA). Gemcitabine (GEM) was brought from Selleckchem. The PBS medium, Annexin V-FITC/PI apoptosis detection kit, and BCA protein assay kit were purchased from KeyGEN Biotech (Jiangsu, China). 5%BSA, paraformaldehyde (PFA), electrophoresis solution, transfer solution, CCK-8 kit, RIPA, and SDS-PAGE gel were all provided by Beyotime Biotech (Beijing, China). The mouse tumor and spleen dissociation kits were obtained from Miltenyi (MACS, Miltenyi Biotec, Germany). Anti-mouse PE-CD11b, anti-mouse FITC-Gr-1, anti-mouse PerCP/Cy5.5-Ly6C, anti-mouse APC-Ly6G, anti-mouse PE NKG2D, anti-mouse APC-CD49b, anti-mouse FITC-CD4, and anti-mouse APC-CD25 were purchased from BioLegend. RT-PCR primers were designed and provided by Sangon Biotech (Shanghai, China). Trizol reagent, Tween-20, and 20× TBS buffer were obtained from Thermo Scientific (Rockford, IL). An ECL Kit was provided by Tanon Biotech (Shanghai, China).
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