Fitc labeled anti igg1

Spleens were isolated from MRL.Fas^lpr mice at 25 weeks of age, and immune cell composition was analyzed by flow cytometry. Spleen cells were stained with the following monoclonal antibodies in 0.5% BSA/PBS at 4°C for 15 min: FITC-labeled anti-B220, APC-labeled anti-CD3, PE-labeled anti-CD4, FITC-labeled anti-CD8, FITC-labeled anti-CD11c, PE-labeled anti-CD11b, APC-labeled anti-CD138, FITC-labeled anti-IgG1 (BD Biosciences, San Jose, CA, USA), FITC-labeled anti-CD4, and PE-labeled anti-Foxp3 (eBioscience). Data were collected using FACSCalibur and analyzed with CellQuest Pro software (BD Biosciences).

A number of 1×10⁵ ATMSC was collected in 200 μL PBS containing 2% FBS. After that, ATMSC were incubated with antibodies for 30 minutes at 4°C, including FITC-labeled anti-CD90 (BioLegend, San Diego, CA; 328107), PE-labeled anti-CD105 (BioLegend; 323206), PE-labeled anti-CD73 (BD Biosciences, San Diego, CA, 550257), FITC-labeled anti-CD31 (BioLegend; 303103), FITC–labeled anti-CD45 (BD Biosciences; 560976) and PE-labeled anti-CD34 (BD Biosciences; 560941). Cells stained with PE-labeled anti-IgG1 (555749; BD Biosciences), and FITC-labeled anti-IgG1 (555748; BD Biosciences) were used as the isotype controls. After that, cells were washed with PBS and resuspended in 300 μL PBS containing 2% FBS. The expression of MSC markers was analyzed by a flow cytometer (BD LSRFortessa X-20; BD Biosciences).