Thermo Scientific Bond-Breaker TCEP Solution, Neutral pH is a stable, 0.5M solution of the thiol-free, phosphine-based TCEP compound, useful as a 10X stock for addition to SDS-PAGE sample loading buffers to reduce protein disulfide bonds.
Features of Bond-Breaker TCEP Solution, Neutral pH:
• Odorless—unlike DTT and β-ME, TCEP is odor-free, contributing to a healthier lab environment • Efficient—5 to 50 mM TCEP thoroughly reduces most peptide or protein disulfide bonds within a few minutes (i.e., just as effective as DTT) • Specific—selective and complete reduction of even the most stable water-soluble alkyl disulfides • Fast—reduces protein disulfides at room temperature and pH 5 in less than five minutes • Stable—resistant to air oxidation; nonvolatile and nonreactive toward other functional groups found in proteins • Versatile—reduces peptides and proteins over a broad range of pH, salt, detergent and temperature conditions • Compatible—removal of the reducing agent is not necessary before most applications, (e.g. histidine-tagged protein purification, maleimide conjugations), because TCEP does not contain sulfhydryl groups
Bond-Breaker TCEP Solution is a potent, odorless, thiol-free reducing agent with broad application to protein and other research involving reduction of disulfide bonds. This product is an effective and convenient replacement for B-mercaptoethanol or DTT in SDS-PAGE sample buffers. The neutral pH of this reagent provides sharp bands and avoids exposing proteins to the strong acid of TCEP·HCl, which can result in hydrolysis and carbohydrate modification.
Considerations for use of Bond-Breaker TCEP Solution: • Reduction occurs over a wide range of pH (pH 4.0-9.0) and temperature (5 to 95°C). • Most proteins are reduced efficiently without a denaturant. However, adding a denaturant such as guanidine·HCl may aid in exposing internal disulfides to the Immobilized TCEP. • Urea is not recommended as a denaturant as it forms cyanates that react with sulfhydryl groups. • Do not allow metals to contact the TCEP solution as this will decrease TCEP activity. • Including 5 to 20 mM EDTA in the sample buffer during reduction helps prevent reoxidation of the sulfhydryl groups by divalent metals such as Zn 2+, Cu 2+ and Mg 2+. • The reduced sample should be used immediately after reduction because disulfides will reform over time.