CST, 5870S, Phosphatase Inhibitor Cocktail (100X)

Buffer for studying in the research area. Product Usage Information 1. Briefly vortex the Phosphatase Inhibitor Cocktail (100X) before use. 2. Just prior to lysing cells: a. Western Blot or Immunoprecipitation Cell Lysis, dilute the cocktail 1:100 in desired lysis buffer to obtain a 1X working concentration. b. PTMScan (R) Cell Lysis, dilute the cocktail 1:50 in urea lysis buffer to obtain a 2X working concentration. This higher concentration is required due to the concentrated nature of the lysate. Storage Store the undiluted 100X cocktail at 4ºC. Do not freeze. Background Dynamic protein phosphorylation is a key cellular signaling mechanism by which a broad spectrum of cellular processes is regulated. In order to study the phosphorylation status of specific target proteins the phosphorylated residue of interest must remain intact. When cells are lysed to make whole cell extracts, a loss of normal cellular signaling regulation occurs, and phosphatases within the cell extract are free to dephosphorylate proteins in an uncontrolled manner. The addition of phosphatase inhibitors to the cell lysis buffer aids in the preservation of phosphorylated residues at the time of cell disruption. Alternate Names aprotinin; beta-glycerophosphate; cocktail; leupeptin; Phosphatase Inhibitor Cocktail; Phosphatase Inhibitors; sodium fluoride; sodium orthovanadate; sodium pyrophosphate; tablet