ab8312 staining human PBMC, huh-7 cells by ELISA.  Cells were incubated in their respective growth medium with 10% fetal calf serum. At the end of the incubation they were washed with PBS (4°C) and the pellet was used to determine the CsA content.  1st step: cyclophilin B (ab53870, 1/1500 carbonate pH 8.5) was bound to the plate (overnight 4°C), 2nd step blocking agent, 3rd step incubation with cell lysate incubated with CsA or analogs (CsA-OH) overnight,4th step ab8312 mouse anti CsA, 1/500 for 2 hours at 37°C 5th step ab6729 (1/2000) Alkalline phosphatase reading at 405 nmSee Abreview