Application

An excellent reagent for maintaining SH groups in reduced state; quantitatively reduces disulfides. DTT is effective in sample buffers for reducing protein disulfide bonds prior to SDS-PAGE. DTT can also be used for reducing the disulfide bridge of the cross-linker N,N′-bis(acryloyl)cystamine to break apart the matrix of a polyacrylamide gel. DTT is less pungent and is less toxic than 2-mercaptoethanol. Typically, a seven fold lower concentration of DTT (100 mM) is needed than is used for 2-mercaptoethanol (5% v/v, 700 mM).

DL-Dithiothreitol is a pivotal component used in the preparation of buffer which in turn is utilized in the platelet cytosol preparation. It is also important in isolation of RNA by using guanidinium salts and homogenisation of tissue and cells.

Biochem/physiol Actions

DL-Dithiothreitol denatures proteins by reducing the disulphide bonds and reducing them to SH groups. It also prevents intramolecular and intermolecular disulfide bond formation between cysteine residues of proteins.