三(2-羧基乙基)膦盐酸盐(TCEP盐酸盐)是一种新型的巯基化DNA的还原剂和去保护剂。巯基化DNA末端硫原子在溶液中趋向于形成二聚体,特别是在存在氧气的情况下。这种二聚化大大降低了一些偶联反应实验(如DNA在生物感应器中的固定)的效率;而在DNA溶液中加入三(2-羧基乙基)膦盐酸盐(TCEP盐酸盐),反应一段时间后除去,就可以降低DNA的二聚化。
TCEP hydrochloride has been introduced which oers the prospect of serving as an alternative to the more commonly employed DTT in the NF-κB-DNA binding reactions in vitro, using recombinant p50 protein and a 32 P-labelled κB oligonucleotide. DTT promotes NF-κB-DNA binding in concentrations from 0.25 to 2.6 mM in binding reactions. However, in the presence of 0.25 mM DTT, inhibition of NF-κB binding is seen only at Hg 2+ concentrations greater than 100 μM and results are highly variable. In contrast, TCEP hydrochloride promotes NF-κB-DNA binding in a dose-related manner in concentrations from 0.25 to 6 mM. In the presence of even 6 mM TCEP hydrochloride, Hg 2+ prevents NF-κB-DNA binding at concentrations as low as 20 μM in binding reactions. The human lactoferrin (hLF) peptide is dissolved in phosphate buffer to a concentration of 0.1 mm. Reduction of the disulfide bonds is obtained by adding a 30-fold molar excess of TCEP hydrochloride with subsequent incubation for 2 h at 37 ℃.