Table of Contents
What is TCEP used for?
TCEP (tris(2-carboxyethyl)phosphine) is a reducing agent frequently used in biochemical and molecular biology applications. TCEP is often used as a reducing agent to break disulfide bonds within and between proteins as a preparatory step for gel electrophoresis.
How long does TCEP last?
TCEP-HCl has been shown to be stable, with 80% of its original reduc- ing ability intact after 21 days at pH values between 1.5 – 11.1. At neutral pH values, phosphate buffered saline and other phosphate containing re- agents can facilitate the oxidation of TCEP-HCl, 50 to 100% oxidation after 72 hours.
How much TCEP should I use?
Efficient—For most applications, 5 to 50 mM TCEP provides sufficient molar excess to effectively reduce peptide or protein disulfide bonds within a few minutes at room temperature.
How do you neutralize TCEP?
(Note: TCEP solutions are commonly neutralized due to having a pH of ~2.5 in water.) Add 5.733 g of TCEP to a 50 ml conical vial, making sure to get as much of the TCEP as possible into the vial. Add 35 ml of cold molecular biology grade water to the vial, and dissolve the TCEP.
Is TCEP toxic?
TCEP is toxic flame retardant added to polyurethane foam and is found in furniture and baby products, as well as some plastics and carpet backing.
What does TCEP do to proteins?
Tris (2-carboxyethyl) phosphine (TCEP) is a reducing reagent used in molecular biology and protein biochemistry research. Researchers often add TCEP to denature proteins during preparation of protein samples for gel electrophoresis.
Is TCEP better than DTT?
For long-term storage of proteins, TCEP is significantly more stable than DTT without metal chelates such as EGTA in the buffer, whereas DTT is more stable if metal chelates are present. Thus TCEP has advantages over DTT, although the choice of reductant is application specific.
Where do you store TCEP?
Following reconstitution, aliquot and freeze (-20°C). TCEP retains stability when stored at an acidic or alkaline pH; avoid storage at pH 7.0-8.0. Also avoid exposure to high concentrations of phosphate ions (>150 mM). Stock solutions are stable for up to 3 months at -20°C.
Is TCEP a flame retardant?
Background on TCEP TCEP (CASRN 115-96-8) is a colorless liquid. The primary use for TCEP is as a flame retardant in paint and coating manufacturing.
What is the difference between DTT and TCEP?
Can TCEP replace DTT?
The newly introduced sulfhydryl reductant tris(2-carboxyethyl)phosphine (TCEP) is a potentially attractive alternative to commonly used dithiothreitol (DTT).
Can TCEP be frozen?
TCEP is also capable of functioning as a reducing agent even in pH range of 1.5-8.5. Is more resistant to air-oxidation as compared to DTT. Following reconstitution, aliquot and freeze (-20°C). TCEP retains stability when stored at an acidic or alkaline pH; avoid storage at pH 7.0-8.0.
How long does TCEP last in buffers?
TCEP is not particularly stable in phosphate buffers, especially at neutral pH. Experiments indicate that TCEP completely oxidizes within 72 hours in 0.35M phosphate-buffered saline (PBS), pH 7.0. Approximately 50% oxidation occurs in the same amount of time in 0.15M PBS, pH 8.0.
Does 280 absorb TCEP?
Moreover, TCEP does not absorb UV light in the range of 250 – 280 nm, making it useful for monitoring reactions with proteins that contain aromatic amino acid residues.
Does TCEP interfere with maleimide?
A number of recent reports, however, have confirmed that TCEP and THPP do indeed react with maleimides to reduce conjugation yields significantly.
What is the role of DTT?
The main role of DTT in molecular biological assays is to keep proteins in a reduced state [3,4]. Thiol containing compounds have, however, also been shown to be very effective at protecting DNA from irradiative damage [5,6,7,8], which is thought to be due to their ability to scavenge oxygen and nitrogen radicals.