Thermo Scientific Pierce TCEP-HCl is pure, crystalline Tris(2-carboxyethyl)phosphine hydrochloride, a thiol-free compound that is highly effective at reducing protein and peptide disulfide bonds, as well as being odorless and stable.
TCEP selectively and completely reduces even the most stable water-soluble alkyl disulfides over a wide pH range. Reductions frequently require less than 5 minutes at room temperature. TCEP is non-volatile, odorless, and unlike most other reducing agents, is resistant to air oxidation. Compared to DTT, TCEP is more stable, more effective, and able to reduce disulfide bonds at lower pHs.
- Odorless – reduce proteins at your desktop; contributes to a healthier lab environment
- Specific – selective and complete reduction of even the most stable water-soluble disulfides
- Simple – effective reduction 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
Advantages of TCEP-HCl over traditional alternatives for reducing disulfides:
- Odorless – Unlike DTT or BME, TCEP is odor-free, so reductions can be carried out conveniently on the bench top.
- Stable in air – The inherent stability of the TCEP moiety eliminates the need for any special precautions to avoid oxidation when handling, using or storing TCEP.
- Efficient – For most applications, 5-50 mM TCEP provides sufficient molar excess to effectively reduce peptide or protein disulfide bonds within a few minutes at room temperature.
- Compatible – With TCEP, removal of the reducing agent is not necessary prior to most applications, (e.g. histidine-tagged protein purification, maleimide conjugations).
The ability of trialkylphosphine compounds to reduce protein disulfide bonds have been known for many years.1,2 Phosphines are stable in aqueous solution, selectively reduce disulfide bonds, and are essentially nonreactive toward other functional groups commonly present in proteins.2 Trialkylphosphines, however, were hindered by their instability in water and their disagreeable odor. These obstacles were overcome by discovery of tris(2-carboxyethyl)phosphine (TCEP).3-17
Reduction of Disulfide bonds using TCEP Solution.
Tris[2-carboxyethyl] phosphine (TCEP) is a popular odorless reducing agent for protein applications. The uses for TCEP have recently been expanded with the introduction of a stable, neutral pH TCEP solution, Bond-Breaker TCEP Solution, that is ideal for preparing proteins for SDS-PAGE analysis. Reducing peptides and proteins without contaminating the sample with reducing agent is now possible with Immobilized TCEP Gel where TCEP has been covalently attached to an agarose support.
|Properties of TCEP reducing agent.
||> 310 g/L (1.08 M)
||Store at room temperature
Considerations for use of TCEP:
- TCEP is generally very soluble in aqueous buffers at nearly any pH. Therefore, working concentrations and 10X stock solutions may be readily prepared in most aqueous buffers
- TCEP is stable in aqueous, acidic, and basic solutions. When TCEP is dissolved directly in water, the resulting pH is approximately 2.5.
- TCEP is not particularly stable in phosphate buffers, especially at neutral pH. Therefore, if TCEP is to be used in PBS buffers, prepare the working solution immediately before use.
- TCEP may be used as a substitute for DTT or 2-mercaptoethanol (2-ME) in sample loading buffer for SDS-PAGE; use a final concentration of 50 mM TCEP.
- Because TCEP is charged in solution, it is not compatible for use in isoelectric focusing.
- Duchardt F. et al. (2009) A cell-penetrating peptide derived from human lactoferrin with conformation-dependent uptake efficiency. J Biol Chem. 284, 36099-108.
- Hou J. M. et al. (2008) Atpase activity of Mycobacterium tuberculosis SecA1 and SecA2 proteins and its importance for SecA2 function in macrophages. J. Bacteriol. 190, 4880-7.
- Feng J. et al. (2008) Quantitative proteomic profiling of muscle type-dependent and age-dependent protein carbonylation in rat skeletal muscle mitochondria. J Gerontol A Biol Sci Med Sci. 63, 1137-52.
- Chan C. T. et al. (2008) Molecular imaging of the efficacy of Heat Shock Protein 90 inhibitors in living subjects. Cancer Res. 68, 216-26.
- Nossal N. G. et al. (2007) Architecture of the bacteriophage T4 replication complex revealed with nanoscale biopointers. J. Biol. Chem. 282, 1098-108.
Immobilized TCEP Disulfide Reducing Gel
Bondbreaker TCEP Solution
Sulfhydryl-reactive Biotinylation Reagents
Sulfhydryl-reactive Crosslinking Reagents
SulfoLink Coupling Gel (Sulfhydryl immobilization)
Maleimide-activated Carrier Proteins