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Gene certain primers and Taq Guy probes for quantitative RT PCR have been made employing Universal Probe Library as comprehensive previously. The Cp values for each HDAC and Sirtuin gene have been normalized to the Cq values from the constitutively expressed ? actin gene. Western blot examination Total proteins from H9c2 cells have been extracted utilizing radio immunoprecipitation buffer in accordance to your makers protocol. The nuclear and cytoplasmic and fractions had been separated using the NE PERTM approach. For western blot examination, equal quantities of protein from each sample have been separated using 10% SDS Page. After electrophoresis, the protein samples had been transferred to Immobilon P membranes working with a Trans Blot elec trophoresis transfer cell.
Several HDACs, sirtuins and MAP kinases were detected on western blots with mono specific main antibodies.
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