A431 cells were grown in a CO2 incubator at 37°C to approximately 70% confluency in DMEM media supplemented with 10% fetal bovine serum. Cells were subsequently treated with 0.66 µM TSA in DMSO (TSA treated cells) or with DMSO only (untreated cells) and returned to the incubator for 5h. Cell lysates were subsequently harvested. TSA treated and untreated lysates are supplied as lyophilized powder.
Figure 1: Western blot of TSA-treated and untreated lysates probed with an anti-acetyl lysine MAb
Legend: Lysates were made from A431 cells that were treated (+) or untreated (-) with the HDAC inhibitor trichostatin A (TSA). Each lane represents 20 µg of cell lysate. Lysates were run on SDS-PAGE and transferred to PVDF membranes. Membranes were probed with anti-acetyl lysine antibody (Cat. # AAC01) at 1:500 dilution in TBST in the presence of 10 µg/ml BSA or 10 µg/ml of Ac-BSA. The Ac-BSA eliminates all acetyl lysine signal on the membrane. Prominent bands in the TSA-treated lanes represent acetyl tubulin (55 KDa) and acetyl histones (14-16 KDa).
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