Useful tips

How much TCEP should I use?

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.

How does TCEP work?

TCEP selectively and completely reduces even the most stable water-soluble alkyl disulfides over a wide pH range. TCEP effectively reduces disulfide bonds over a broad pH range. TCEP is resistant to air oxidation. Compared to DTT, TCEP is more stable, more effective, and able to reduce disulfide bonds at lower pHs.

How do you store TCEP powder?

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 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.

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.

What can I use instead of DTT?

Yes you can use beta-mercaptoethanol as an alternative to DTT if you don’t have DTT. Generally DTT is preferred because it is not volatile and therefore doesn’t smell nearly as much as beta-mercaptoethanol, is effective at lower concentrations and is more stable. You could also use TCEP.

Does TCEP work at low pH?

TCEP-HCl is an odor- less (non-volatile) reducing agent that has been found to be more stable and effective than dithiothreitol (DTT) and able to work well at lower pH levels. TCEP-HCl is stable in aqueous solutions and have been found to be in effect unreactive toward other functional groups in proteins.

How do you make a 0.5 M TCEP?

Add 35 ml of cold molecular biology grade water to the vial, and dissolve the TCEP. This resulting solution is very acidic, with an approximate pH of 2.5. Bring the solution to pH 7.0 with 10 N NaOH or 10 N KOH.

What is chlorinated Tris?

Chlorinated tris is a chemical flame retardant. Adding flame retardants to products is one way to reduce flammability. Chlorinated tris can be gradually released from treated products into indoor environments, including houses, schools, day care centers, offices, and cars.

How do you dispose of DTT?

Place spilled material in a designated, labeled waste container. Dispose of via a licensed waste disposal contractor. For emergency responders : If specialised clothing is required to deal with the spillage, take note of any information in Section 8 on suitable and unsuitable materials.

Is TCEP better than DTT?

TCEP HCl has the advantages of being odorless, more powerful, irreversible, more hydrophilic, and more resistant to oxidation in air. DTT is a protective agent for reducing S-S TO SH groups. Used as a strong reducing agent for proteins and enzymes.

How big is the molecular weight of TCEP?

It is often prepared and used as a hydrochloride salt (TCEP-HCl) with a molecular weight of 286.65 gram/mol. It is soluble in water and available as a stabilized solution at neutral pH and immobilized onto an agarose support to facilitate removal of the reducing agent.

What kind of reducing agent is TCEP used for?

TCEP (tris(2-carboxyethyl)phosphine) is a reducing agent frequently used in biochemistry and molecular biology applications.

What can Thermo Scientific TCEP be used for?

The Thermo Scientific™ TCEP is a potent, versatile, odorless, thiol-free reducing agent with broad application to protein and other research involving reduction of disulfide bonds (Figure 1). The unique compound is easily soluble and very stable in many aqueous solutions.

How is TCEP used in the labeling of cysteines?

TCEP is particularly useful when labeling cysteine residues with maleimides. TCEP can keep the cysteines from forming di-sulfide bonds and unlike dithiothreitol and β-mercaptoethanol, it will not react as readily with the maleimide. However, TCEP has been reported to react with maleimide under certain conditions.