TCEP HCl (Tris(2-carboxyethyl)phosphine HCl) is a non-thiol reducing agent that is more stable than other chemical reducing agents and produces faster SS reduction reactions. TCEP HCl is a trialkylphosphine that selectively reduces protein breakdown without changing its properties or interacting with thiol directing agents in the reaction mixture. TCEP HCl is also a extensively usedreducing agent in DNA/AuNP chemistry.

Physicochemical Properties

Molecular Formula	C9H16CLO6P
Molecular Weight	286.65
Exact Mass	286.037
CAS #	51805-45-9
Related CAS #	TCEP-d16 hydrochloride;1174025-33-2
PubChem CID	2734570
Appearance	White to light yellow solid powder
Density	1.041 g/mL at 25 °C
Boiling Point	519.4ºC at 760 mmHg
Melting Point	177 °C
Flash Point	267.9ºC
Index of Refraction	n20/D 1.367
LogP	1.694
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	9
Heavy Atom Count	17
Complexity	222
Defined Atom Stereocenter Count	0
InChi Key	PBVAJRFEEOIAGW-UHFFFAOYSA-N
InChi Code	InChI=1S/C9H15O6P.ClH/c10-7(11)1-4-16(5-2-8(12)13)6-3-9(14)15;/h1-6H2,(H,10,11)(H,12,13)(H,14,15);1H
Chemical Name	3-[bis(2-carboxyethyl)phosphanyl]propanoic acid;hydrochloride
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.(2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

The introduction of TCEP hydrochloride presents a potential substitute for the widely used DTT in vitro NF-κB-DNA binding processes. This can be achieved through the use of a 32P-labelled κB oligonucleotide and recombinant p50 protein. In binding experiments, DTT increases NF-κB-DNA binding at doses ranging from 0.25 to 2.6 mM. Nevertheless, only at Hg2+ concentrations higher than 100 μM can suppression of NF-κB binding occur in the presence of 0.25 mM DTT, and the outcomes vary greatly. On the other hand, at doses ranging from 0.25 to 6 mM, TCEP hydrochloride enhances NF-κB-DNA binding in a dose-dependent manner. Hg2+ inhibits NF-κB-DNA binding in binding assays at concentrations as low as 20 μM when even 6 mM TCEP hydrochloride is present[1]. Phosphate buffer is used to dissolve the human lactoferrin (hLF) peptide to a concentration of 0.1 mm. By adding a 30-fold molar excess of TCEP hydrochloride and then incubating for two hours at 37 °C, the disulfide linkages are reduced[2].

References

[1]. 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]. A cell-penetrating peptide derived from human lactoferrin with conformation-dependent uptake efficiency. J Biol Chem. 2009 Dec 25;284(52):36099-108. [3]. 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]. 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. [5]. Han JC, Han GY. A procedure for quantitative determination of tris(2-carboxyethyl)phosphine, an odorless reducing agent more stable and effective than dithiothreitol. Anal Biochem. 1994;220(1):5-10.

Solubility Data

Solubility (In Vitro)	DMSO: 100 mg/mL (348.86 mM) H2O: 50 mg/mL (174.43 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 100 mg/mL (348.86 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.4886 mL	17.4429 mL	34.8857 mL
	5 mM	0.6977 mL	3.4886 mL	6.9771 mL
	10 mM	0.3489 mL	1.7443 mL	3.4886 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.