Ishikawa cells (human endometrial adenocarcinoma cell line) were cultured according to standard protocol. The culture medium was aspirated and cells rinsed with Ca and Mg free HBSS. Cells were treated with 0.25% Trypsin and incubated at 37° for 5 minutes. Cells were aspirated and pelleted at 900 x g for 5 minutes. Cells were washed twice with PBS. The cells were then fixed with 4% paraformaldehyde in PBS for 15 minutes at room temperature. The cells were then washed as stated previously. Permeabilization and blocking was performed by incubating in 5% BSA and 0.1% Triton-X in PBS for 20 minutes at room temperature. The cells were then washed as previously stated. The primary antibody for OXTR (Product # PA5-19038) was used at a 1:200 dilution in a 5% BSA, PBS solution and incubated for 120 minutes at room temperature. The cells were washed as stated previously. The secondary Alexa 488 antibody (Product # A-11055) was used at a 1:2000 dilution in 5% BSA, PBS and incubated in the dark for 45 minutes. The cells were washed and resuspended in PBS and analyzed through flow cytometry. Data courtesy of the Antibody Data Exchange Program.