Analysis Note

Bio-testsGel has an effective TCEP concentration ≥8 µmol/mL gel

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).