By Sumitra Deb, Swati Palit Deb
An fundamental number of novel suggestions that experience confirmed most beneficial for learning the physiological homes of p53 either in vitro and in vivo. The strategies supply confirmed recommendations to difficulties in learning the purification, objective id, gene expression, quantitation, interplay, signaling, transactivation, and transrepression of p53. The tools also are worthy for delineating the features of alternative proteins that can act as tumor or progress suppressors. each one strategy contains step by step directions, troubleshooting notes, a theoretical overview, and dialogue of linked difficulties that will come up in the course of the process research.
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3. Hybridization buffer: 5X SSC, 50% formamide, 5X Denhardt’s solution, 1% sodium dodecyl sulfate (SDS), and fresh 100 µg/mL heat-denatured sheared nonhomologous salmon sperm DNA (store at –20°C). 4. HotPrime™ DNA labeling kit (GenHunter). 3. 1. RNA Isolation from Cell Cultures Total RNA can be isolated with one-step acid-phenol extraction method by using RNApure™ Reagent. 1. After removal of the cell culture medium and wash step with 10–20 mL of cold phosphate-buffered saline (PBS), set the flask or plate on ice.
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