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 light sensitive?

Note: Cover the tubes with aluminum foil, because TCEP is light sensitive. Note: Stock solutions are stable for 3 months at -20°C.

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 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.

Does TCEP react with maleimide?

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. TCEP is also used in the tissue homogenization process for RNA isolation.

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.

What other uses does chlorinated Tris have?

Chlorinated tris is a chemical flame retardant. Adding flame retardants to furniture products is one way to reduce flammability. Polyurethane foam treated with chlorinated tris may be used in upholstered furniture. Textiles and plastic treated with chlorinated tris may be used in furniture products.

What chemicals make up chlorinated Tris?

Chlorinated Tris, or Tris (1,3-dichloro-2-propyl) phosphate is a chlorinated phosphate ester (#1). It is a colorless, viscous liquid at temperatures greater than 27 degrees C (#4). Chlorinated Tris in an additive flame retardant, meaning that is not chemically bound to products and can escape over time.

What’s the difference between DTT and TCEP reaction?

TCEP reaction is irreversible, whereas DTT reaction is reversible. Compared to DTT, TCEP is preferred for labeling cysteine residues with maleimides. DTT contains thiols and shows reactivity with maleimides, which reduces the labeling efficiency.

Do you need to remove TCEP from the labeling reaction?

Note: TCEP does not contain thiols so there is no need to remove it from the labeling reaction. But TCEP may react with maleimides under certain conditions, such as acidic conditions, at 20°C, and too much TCEP for labeling of proteomic samples.

What is the use of TCEP in protein preparation?

Researchers often add TCEP to denature proteins during preparation of protein samples for gel electrophoresis. TCEP has also been useful for long-term storage of protein and preparation of samples in many other applications, such as:

What is the protocol for TCEP and maleimide reaction?

Protocol for tcep and maleimide reaction? I am trying to see if my protein has irreversible, reversible, or it forms disulfide bonds. Previously, I mixed my protein ( 25uM) with MAL-PEG 5000 ( 250 uM) to see if the maleimide will react with the protein, It did not react with the protein, so the molecular weight did not go up.