Lck

1ml per sample. The samples were incubated at 4⁰C overnight. The following day, the samples were washed with DPBS at 4⁰C and then washed with 2ml 1 perm buffer twice. Anti-mouse Foxp3- PE Ab was added in a 1:100 dilution for 30 mins at 4⁰C. The samples were washed with perm buffer and resuspend in DPBS for flow cytometry analysis. Table 2.4 Antibody panel for SHPS-1 Phenotyping Panel PerCP FITC APC PE PB DC 1 CD45 CD11b CD11c CD103 B220 DC 2 CD4 CD11b CD11c CD8 B220 Treg CD3 CD25 FoxP3 CD4 2.2.1.2.5 Analysis of Cell Populations in Allo- and Xeno-transplantation Recipients The spleen, ALN and DLN were harvested from mice at certain time points post allo or xeno-transplantations. SHPS-1 and WT mice who were the recipients of skin allo-transplants, DEREG, WT and Rag-/- mice who were the recipients of NICC xeno-transplants were harvested for flow cytometry analysis. The procedure for isolation of splenocytes is described in 2.2.1.1.2, then staining protocols were followed as per the same steps as 2.2.1.2.4. Splenocytes and lymphocytes were stained with anti-mouse-CD4-PerCP, anti-mouse-CD3-FITC, anti-mouse-CD25-APC, anti-mouse-CD8-PE, anti-mouse-CD4-PB, anti-mouse-B220-PB, and anti-mouse-CD3-PB. The panels were as shown in Table 2.5. Table 2.5 Antibody panel for Analysis of Cell Populations in Allo- and Xeno-transplantation mouse recipient Panel PerCP FITC APC PE PB DN T Cells CD4 CD3 CD25 CD8 B220 Treg 1 CD3 CD25 FoxP3 CD4 Treg 2 CD4 GFP CD25 FoxP3 CD3 For each…