Anti-Retinoid X Receptor Alpha Antibody [EPR7106]

HL-60 cells are able to differentiate to neutrophils after all trans retinoic acid (RA) treatment. RXRalpha expression was assayed using Anti-Retinoid X Receptor alpha antibody [EPR7106] (ab125001). The purpose of the present experiment was to establish the experimental conditions, as an antibody that we previously used- ab118329- is no longer available. Cell culture volumes containing two million cells, RA treated, were harvested, fixed for 10 min with 2% paraformaldehyde, and permeabilized with ice cold methanol. After 3 washes with PBS, the cells were incubated with Anti-Retinoid X Receptor alpha antibody [EPR7106] (ab125001) that we previously labeled with the ThermoFisher Alexa Fluor® 488 dye. We used 2.5µl labeled antibody/200 µl PBS. Incubation was overnight at 4C on a rocker in the dark. This was followed by 3 washes with 500µl PBS-0.1%Tween and resuspension in 200 µl PBS before reading. The result was comparable to what we used to obtain with the ab118329. The image shows the histograms for unlabeled (background signal) and RXRα- Alexa488 labeled cells.