52486
Tris(2-carboxyethyl)phosphine, immobilized on Agarose CL-4B
Synonym(s):
TCEP-Agarose, Tris(2-carboxyethyl)phosphine-Agarose
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About This Item
UNSPSC Code:
12352204
NACRES:
NA.25
Recommended Products
storage temp.
2-8°C
Application
Tris(2-carboxyethyl)phosphine, immobilized on agarose CL-4B may be used in preliminary off-line experiments to study its ability to cleave disulfide bonds by both under flow conditions using sequential injection analysis (SI). It may also be used as a reducing agent in bio-tests for disrupting the disulfide bonds within and between the proteins.
Biochem/physiol Actions
Tris(2-carboxyethyl)phosphine also referred to as TCEP reduces the disulfides by the usual mechanism of reduction of disulfides by phosphines in water. Tris(2-carboxyethyl)phosphine, immobilized on Agarose CL-4B,Trialkylphosphines (TCEP) are highly effective agents for reducing disulfide bonds in proteins, peptides and other disulfide bondcontaining molecules and are relatively non-reactive toward other functional groups. The trialkylphosphine TCEP was first described by Levison et al. as an odorless and efficient reductant of alkyl disulfides over a wide pH range. TCEP is stable in aqueous solutions and does not undergo the rapid oxidation that often occurs with other reducing agents such as dithiotreitol (DTT) and β-mercaptoethanol (BME, 2-ME). TCEP does not interfere with commonly used sulfhydryl-reactive reagents (e.g., maleimide crosslinkers). Nevertheless, many protocols require recovery of the reduced sample separate from the reducing agent. Our Tris(2-carboxyethyl)phosphine, immobilized on Agarose CL-4B eliminates the need to use laborious and troublesome gel filtration methods to separate the reduced sample from the reducing agent. Immobilized TCEP Disulfide Reducing Gel may be adapted conveniently to a variety of scales and formats. Examples are given for batch, spin cup column and gravity-flow column procedures. For small-scale reductions, the most complete sample recovery is made using the spin-cup column procedure (see Related Thermo Scientific Products Section).
Analysis Note
Bio-tests
Gel has an effective TCEP concentration ≥8 μmol/mL gel
Gel has an effective TCEP concentration ≥8 μmol/mL gel
Storage Class Code
10 - Combustible liquids
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Certificates of Analysis (COA)
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On-line cleavage of disulfide bonds by soluble and immobilized tris-(2-carboxyethyl)phosphine using sequential injection analysis
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J C Han et al.
Analytical biochemistry, 220(1), 5-10 (1994-07-01)
The concentration of tris(2-carboxyethyl)phosphine (TCEP) can be conveniently determined by measuring the amount of 2-nitro-5-thiobenzoate (NTB) formed after reaction with 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB). This method utilizes the fact that TCEP reduces DTNB rapidly and stoichiometrically to generate two equivalents of
Burns, J.A.
The Journal of Organic Chemistry, 56, 2648-2648 (1991)
Reductive cleavage of cystine disulfides with tributylphosphine.
U T Rüegg et al.
Methods in enzymology, 47, 111-116 (1977-01-01)
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