Nuclear Transcription Factor Detection

Cellular fate is determined by the activity of nuclear transcription factors. Here, we describe a series of protocols for detecting transcription factors at both the transcript and protein levels in human adipose cells. Methods for analysis of transcript include RNA extraction, reverse transcription polymerase chain reaction (RT-PCR), endpoint PCR, and RT-qPCR. Evaluation of protein expression includes protocols for protein extraction, quantification by Bradford assay, SDS-PAGE, western blotting, and quantification with ImageJ. Each of these steps is critical for a reliable and reproducible assessment of transcription expression and characterization of human adult-derived adipose stromal/stem cells.