Description
TCEP-HCl (Tris(2-carboxyethyl)phosphine hydrochloride) is a high-purity, thiol-free reducing agent widely used in proteomics, RNA , and bioconjugation protocols. It remains active in acidic and denaturing environments, making it ideal for protein sample preparation, disulfide bond reduction, and thiol-specific labeling.
GoldBio’s ≥98% pure TCEP-HCl offers superior stability, odor-free handling, and compatibility with mass spectrometry and oligonucleotide preservation. With unmatched performance in reduction workflows and conjugation reactions, TCEP-HCl is the gold standard for life science researchers requiring precision and reliability.
TCEP 50 g and 100 g ships as multiple 25 g units to qualify for a small quantity exception to hazardous fees.
Due to hazardous goods restrictions, this product cannot ship by ground shipping to Canada or Mexico.
Common Research Applications
(Click each for more information)
Protein Disulfide Bond Reduction in Proteomics
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Purpose: Facilitates protein unfolding by reducing disulfide bonds before SDS-PAGE, 2D gel electrophoresis, or LC-MS/MS.
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How It Works: TCEP-HCl is a thiol-free phosphine reductant that cleanly cleaves disulfide bonds across acidic and denaturing environments.
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Applications: Protein extraction, sample preparation for mass spectrometry, electrophoretic protein separation.
Getz, E. B., Xiao, M., Chakrabarty, T., Cooke, R., & Selvin, P. R. (1999). A comparison between the sulfhydryl reductants tris(2-carboxyethyl)phosphine and dithiothreitol for use in protein biochemistry. Analytical Biochemistry, 273(1), 73–80.
Thiol–Maleimide Conjugation in Bioconjugation Workflows
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Purpose: Maintains cysteine thiols in a reduced state for reaction with maleimide-functionalized probes or drugs.
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How It Works: TCEP prevents thiol oxidation and does not interfere with maleimide chemistry, improving conjugation specificity.
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Applications: Peptide labeling, antibody-drug conjugates (ADCs), biosensors, fluorescent probe conjugation.
Stefano, J. E., Busch, M., Hou, L., Park, A., & Gianolio, D. A. (2013). Micro- and mid-scale maleimide-based conjugation of cytotoxic drugs to antibody hinge-region thiols for tumor targeting. In L. Ducry (Ed.), Antibody–Drug Conjugates (Vol. 1045, pp. 117–127). Humana Press.
Mass Spectrometry Sample Preparation (Reduction–Alkylation)
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Purpose: Ensures full cysteine reduction before digestion to maximize peptide yield and reduce sample variability.
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How It Works: TCEP remains stable in acidic or chaotropic buffers and does not interfere with MS ionization.
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Applications: Bottom-up proteomics workflows, in-gel and in-solution digestion protocols.
Hale, J. E., Butler, J. P., & Gelfanova, V. (2004). A simplified procedure for the reduction and alkylation of cysteine residues in proteins prior to proteolytic digestion and mass spectral analysis. Analytical Biochemistry, 333(1), 174–181.
Reduction in Acidic or Denaturing Buffer Conditions
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Purpose: Enables effective reduction in harsh buffers (e.g., low pH or high concentrations of urea or guanidine) where DTT is unstable.
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How It Works: TCEP is resistant to oxidation and retains reducing power across pH 1.5–9.0, even in chaotropic environments.
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Applications: Membrane protein solubilization, protein unfolding, crystallography buffer systems.
Han, J. C., & Han, G. Y. (1994). A procedure for quantitative determination of tris(2-carboxyethyl)phosphine, an odorless reducing agent more stable and effective than dithiothreitol. Analytical Biochemistry, 220(1), 5–10.
Preservation of Thiol Integrity During RNA or Oligonucleotide Protocols
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Purpose: Maintains the reduced state of thiol-modified oligonucleotides and prevents oxidative RNA degradation.
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How It Works: TCEP protects against disulfide bond formation without interfering with enzymatic reactions such as reverse transcription.
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Applications: Thiol-labeled RNA/DNA probes, transcriptomic workflows, nucleic acid modification.
Rhee, S. S., & Burke, D. H. (2004). Tris(2-carboxyethyl)phosphine stabilization of RNA: Comparison with dithiothreitol for use with nucleic acid and thiophosphoryl chemistry. Analytical Biochemistry, 325(1), 137–143.
Benefits
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TCEP-HCl is stable in air.
- TCEP-HCl is odorless so reactions can be carried out on the bench instead of the fume hood.
- TCEP-HCl is efficient and can be used to reduce disulfide bonds at low concentrations.
- TCEP-HCl is selective for disulfides and does not react with other protein side chains.
- TCEP-HCl may be used as a substitute for DTT at a final concentration of 50mM.
Storage/Handling
Store desiccated at -20°C.
Working Concentration
TCEP is commonly stored in a 0.5M stock solution for up to 3 months at -20°C.
Working concentrations are typically around 5-50mM and should be remade daily.
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