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 32P-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 Hg2+ 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, Hg2+ prevents NF-κB-DNA binding at concentrations as low as 20 μM in binding reactions[1]. 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 ℃[2].
Kinase Assay
Cell Assay
Animal Administration
References
[1]. Dieguez-Acuña FJ, et al. Inhibition of NF-kappaB-DNA binding by mercuric ion: utility of the non-thiol reductant, tris(2-carboxyethyl)phosphine hydrochloride (TCEP), on detection of impaired NF-kappaB-DNA binding by thiol-directed agents. Toxicol In Vitro. 2000 Feb;14(1):7-16.
[2]. Duchardt F, et al. A cell-penetrating peptide derived from human lactoferrin with conformation-dependent uptake efficiency. J Biol Chem. 2009 Dec 25;284(52):36099-108.
[3]. Sequeira MA, et al. Modulating amyloid fibrillation in a minimalist model peptide by intermolecular disulfide chemical reduction. Phys Chem Chem Phys. 2019 Jun 5;21(22):11916-11923.
[4]. Wu R, et al. Effects of Small Molecules on DNA Adsorption by Gold Nanoparticles and a Case Study of Tris(2-carboxyethyl)phosphine (TCEP). Langmuir. 2019 Oct 15;35(41):13461-13468.
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