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Radioimmunoprecipitation assay buffer (RIPA buffer) is a lysis buffer used to lyse cells and tissue for the radio immunoprecipitation assay (RIPA). [1] [2] This buffer is more denaturing than NP-40 or Triton X-100 because it contains the ionic detergents SDS and sodium deoxycholate as active constituents and is particularly useful for disruption of nuclear membranes in the preparation of ...
McIlvaine buffer is a buffer solution composed of citric acid and disodium hydrogen phosphate, also known as citrate-phosphate buffer. It was introduced in 1921 by the United States agronomist Theodore Clinton McIlvaine (1875–1959) from West Virginia University , and it can be prepared in pH 2.2 to 8 by mixing two stock solutions.
Buffer capacity falls to 33% of the maximum value at pH = pK a ± 1, to 10% at pH = pK a ± 1.5 and to 1% at pH = pK a ± 2. For this reason the most useful range is approximately pK a ± 1. When choosing a buffer for use at a specific pH, it should have a pK a value as close as possible to that pH. [2]
Add 2.84 mM of HCl to shift the buffer to 7.3 mM HPO 4 2− and 4.6 mM H 2 PO 4 − for a final pH of 7.4 and a Cl − concentration of 142 mM. The pH of PBS is ~7.4. When making buffer solutions, it is good practice to always measure the pH directly using a pH meter. If necessary, pH can be adjusted using hydrochloric acid or sodium hydroxide.
The buffer can be stored without vanadate at 4 °C for up to 1 year. [10] RIPA buffer releases proteins from cells as well as disrupts most weak interactions between proteins. [9] Recipe: [10] 1% (w/w) Nonidet P-40 (NP-40) 1% (w/v) sodium deoxycholate; 0.1% (w/v) SDS; 0.15 M NaCl; 0.01 M sodium phosphate, pH 7.2; 2 mM EDTA; 50 mM sodium ...
All buffering agents achieve their function because they contain an acidic group (acetate, phosphate, sulphonate ..) or a basic group (amino, pyridyl ..). A consequence of this is that they can form complexes with the biologically important ions Na + , K + , Mg 2+ and Ca 2+ and can compete for the metal ion contained in a metalloprotein .
Composition per 1 mL (once preparation complete): sodium chloride 7.14 mg (122.17 mmol), potassium chloride 0.38 mg (5.097 mmol), calcium chloride dihydrate 0.154 mg (1.04754 mmol), magnesium chloride hexahydrate 0.2 mg (0.983767 mmol), dibasic sodium phosphate 0.42 mg (2.95858 mmol), sodium bicarbonate 2.1 mg (24.998 mmol), dextrose 0.92 mg (5 ...
TCEP is available from various chemical suppliers as the hydrochloride salt. When dissolved in water, TCEP-HCl is acidic. A reported preparation is a 0.5 M TCEP-HCl aqueous stock solution that is pH adjusted to near-neutral pH and stored frozen at -20˚C. [12] TCEP is reportedly less stable in phosphate buffers. [12]