| DEPC is a strong nuclease inhibitor, utilizing a mechanism which modifies His and Tyr residues in proteins. Used as a RNase and DNase inhibitor for Molecular Biology solutions. DEPC is a chemical use to inactivate RNase enzymes and is sensitive to moisture and pH. It decomposes at 155C, to ethanol and carbon dioxide in aqueous solution. DEPC is also sensitive to ammonia, which causes decomposition to urethane, a possible carcinogen. Biochem and Physiological Actions: Inactivates RNase in solution at about 0.1% (v/v), thus protecting RNA against degradation. DEPC is a potent nuclease inhibitor, a 0.1% solution of DEPC is used to inactive RNase. It inhibits the ryanodine binding to ryanodine/Ca2+ receptor channels in skeletal muscle in a dose and time dependent manner and increases the Ca2+ permeability of SR vesicles. Purity: 97% Appearance: Colorless, clear liquid Refractive Index (n20/D): 1.398 (lit.) Boiling Point: 93-94C/18mmHg (lit.) Density: 1.101g/mL at 25C (lit.) Molarity: 6.9M Protocol (Water treatment with DEPC to eliminate RNase): Add 0.1% diethylpyrocarbonate (DEPC) to water, mix overnight, and then autoclave 20min to destroy DEPC by causing hydrolysis of DEPC. Preparation Instructions: DEPC is tested for miscility in 95% ethanol, 1.6ml in 4ml ethanol gives a clear, colorless solution. It is soluble in alcohols, esters, ketones and other hydrocarbons, but only soluble in water to about 0.1%. DEPC hydrolyzes in aqueous solution. In phosphate buffer at 25C, its half-life is 4 minutes at pH6, 9 minutes at pH 7. Hydrolysis is accelerated by TRIS buffer, at 25C, its half-life was reported to be 1.25 minutes at pH 7.5, 0.37 minutes at pH 8.2. Storage and Stability: Diethyl pyrocarbonate is very sensitive to moisture, and as such, is packaged under argon to help reduce exposure to moisture. This product has a shelf-life of 12 months. If the product is exposed to even traces of moisture, it does hydrolyze somewhat. The carbon dioxide which is formed is more soluble in DEPC at 0-4C than at room temperature, and as the product is brought to room temperature before opening, the DEPC at 0-4C can become supersaturated with respect to any dissolved gas. It is advised that the user be familiar with the hazardous nature of DEPC, as described on the Material Safety Data Sheet (MSDS). After opening the bottle for the first time and each time thereafter, layer nitrogen or argon gas over the DEPC and store the closed bottle at 2-8C for optimal stability. It may be helpful to store the bottle inside a sealed plastic bag with desiccant, but with the bottle cap slightly loose. If the bag inflates at all, this is an indication of some degree of decomposition with possible pressure build-up. Once opened, the bottle should not be kept for more than 9 months. Caution: DEPC hydrolyzes to carbon dioxide and ethanol in the presence of moisture, causing increased pressure. Open and use under a fume hood. Keep tightly closed at all times, avoiding moisture. Periodically vent bottle slowly under fume hood. Color will gradually revert from clear to yellow in time. DEPC is a suspected carcinogen. |
