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2-Mercaptoethanol is often used interchangeably with dithiothreitol (DTT) or the odorless tris(2-carboxyethyl)phosphine (TCEP) in biological applications. Although 2-mercaptoethanol has a higher volatility than DTT, it is more stable: 2-mercaptoethanol's half-life is more than 100 hours at pH 6.5 and 4 hours at pH 8.5; DTT's half-life is 40 ...
TCEP is often used as a reducing agent to break disulfide bonds within and between proteins as a preparatory step for gel electrophoresis.. Compared to the other two most common agents used for this purpose (dithiothreitol and β-mercaptoethanol), TCEP has the advantages of being odorless, a more powerful reducing agent, an irreversible reducing agent (in the sense that TCEP does not ...
(2S)-2-Amino-1,4-dimercaptobutane (dithiobutylamine or DTBA), a related dithiol reducing agent, somewhat overcomes this limitation of DTT. [5] Tris(2-carboxyethyl)phosphine ( TCEP ) is an alternative reducing agent that is more stable and effective at low pH, but it is bulky and reduces cystines in folded proteins only slowly.
The thiol reagents are used in excess to drive the equilibrium to the right: + + The reductant tris(2-carboxyethyl)phosphine (TCEP) is useful, beside being odorless compared to β-ME and DTT, because it is selective, working at both alkaline and acidic conditions (unlike DTT), is more hydrophilic and more resistant to oxidation in air ...
The reduction potential of DTE is the same as for DTT, about –0.331 mV. [2] The pK a values of the thiol groups of DTE are 9.0 and 9.9, which is higher than the corresponding values for DTT (9.3 and 9.5). [2] Since reduction of disulfide bonds requires thiolate (ionized thiol), DTE is less efficient at lower pH compared to DTT. [2]
A common 1,4-dithiol is dithiothreitol (DTT), HSCH 2 CH(OH)CH(OH)CH 2 SH, sometimes called Cleland's reagent, for to reduce protein disulfide bonds. Oxidation of DTT results a stable six-membered heterocyclic ring with an internal disulfide bond. Reduction of a typical disulfide bond by DTT via two sequential thiol-disulfide exchange reactions.
[2] Eagle developed his earlier "Basal Medium Eagle" (BME) in 1955–1957 on mouse L cells [3] and human HeLa cells, [4] with 13 essential amino acids and 9 vitamins added. BME contains biotin (vitamin B 7), which Eagle later found to be superfluous. His 1959 "minimal essential medium" doubles the amount of many amino acids to "conform more ...
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 ...