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Wherever possible, you should prepare and use solutions on the same day. However, if you need to make up stock solutions in advance, we recommend that you store the solution as aliquots in tightly sealed vials at -20°C. Generally, these will be useable for up to one month. Before use, and prior to opening the vial we recommend that you allow your product to equilibrate to room temperature for at least 1 hour.
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MALME-3M cells were incubated at 37°C for 24h with vehicle control (0 µM) and 1 µM andrographolide (ab120636). Increased expression of p53 in MALME-3M cells correlates with an increase in andrographolide concentration, as described in literature.
Whole cell lysates were prepared with RIPA buffer (containing protease inhibitors and sodium orthovanadate), 10 µg of each were loaded on the gel and the WB was run under reducing conditions. After transfer the membrane was blocked for an hour using 5% BSA before being incubated with ab26 at 5 µg/ml and ab8227 at 1 µg/ml overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP (ab97040 ) at 1/10000 dilution and visualised using ECL development solution.
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