DL-1,4-Dithiothreitol（578517）reduction and Application

Reduction

DTT is a strong reducing agent, the reduction is largely due to its oxidation statesix membered ring (key two sulfur content) the conformational stability. The redox potential at pH 7 to -0.33 volts. Two of dithiothreitol on a typical two sulfur

Bond reduction is composed of two consecutive steps of thiol disulfide exchange reactions. Two:

Among them, the middle states formed by the first step of the reaction is not stable, because on the DTT second sulfhydryl groups tend to be connected withoxidation of the sulfur atom, the intermediate state quickly converted into cyclicoxidation structure of DTT, thus completing the reduction of two disulfide. Effect of reducing capacity of DTT by the pH value, more than 7 conditions play areduction value only in pH. This is because the thiolate anion only removeprotons (-S -) with reaction activity, thiol (-SH) is not; and the thiol pKa ~ 8.3.

Application

This dimerization greatly reduces the number of coupling reaction (such as DNAin biological sensor fixed) efficiency; while the addition of DTT in DNA solution,reaction after a period of time to remove, can reduce the dimerization of DNA.DTT is also often used for the reduction of two disulfide bonds in proteins, and can be used for protein molecules formed between the stop of cysteines in proteins or molecules of two disulfide. But DTT is often unable to restore theembedded in the protein structure (solvent inaccessible) two disulfide bonds,reducing the two disulfide bonds often need to protein denaturation (high temperature heating or adding denaturants, such as urea or guanidine hydrochloride, 6M 8M 1% SDS).

But DTT is often unable to restore the embedded in the protein structure (solventinaccessible) two disulfide bonds, reducing the two disulfide bonds often need toprotein denaturation (high temperature heating or adding denaturants, such asurea or guanidine hydrochloride, 6M 8M 1% SDS). On the other hand, according to the presence of DTT, two disulfide bond reduction rate is different, you can determine the depth of embedment. Because it is easy to be oxidized in air, so the poor stability of DTT; but the cryopreservation or inert gas treatment canprolong its service life.

Because it is easy to be oxidized in air, so the poor stability of DTT; but thecryopreservation or inert gas treatment can prolong its service life. Due to the protonated sulfur dear nuclear is low, with the decreasing of pH, the effectivereduction of DTT also decreased; while Tris (2-carboxyethyl) phosphine HCl(TCEP hydrochloride) can be used as a low pH values under the conditions ofDTT alternatives, but also is more stable than the DTT.