DESCRIPTION
Molecular Biology Grade, PROTEASE FREE!!
Cleland's reagent, also known as dithiothreitol or DTT is a water soluble protective reagent for sulfhydryl groups. It is capable of maintaining monothiols completely in the reduced state and reduces disulfudes quantitatively. Since its introduction in 1964 by Professor Cleland it has gained widespread acceptance as the reagent of choice for the protection of sulfhydryl groups. DTT readily permeates cell membranes. therefore DTT protects protein sulfhydryls allowing enzyme activity to be restored due to the oxidation of sulfhydryl groups in vitro. 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).
STOCK SOLUTION
Dissolve 3.09 g of DTT in 20 ml of 0.01 M sodium acetate (pH 5.2) and sterilize by filtration through 0.22 ¥ìm filter. Dispense into 1-ml aliquots and store at -20¡É. Under these conditions, DTT is stable to oxidation by air.
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CHEMICAL STRUCTURE : C4 H10 O2 S2
CAS RN : [27565-41-9]
MOLECULAR WEIGHT: 154.25
MELTING POINT: 39-43¡É
CAUTION
HARMFUL IF SWALLOWED OR INHALED. MAY BE HARMFUL IF ABSORBED THROUGH SKIN. CAUSES IRRITATION TO SKIN, EYES AND RESPIRATORY TRACT. MAY AFFECT CENTRAL NERVOUS SYSTEM.
STORAGE CONDITIONS
2-8¡É desiccated / warm to room temperature before opening. Keep dry, protect from light.
REFERENCES 1. Cleland, W.W., Biochemistry 3, 480 (1964) 2. Ruegg, U.T., Rudinger, J., Cleavage of disulfide bonds in proteins Meth. Enzymol. 47, 111 (1977) 3. D.W. Sears, In the investigation of incremental and kinetic reduction in proteins Biochemistry 16, 2031 (1977) 4. McDonald, R.J., Isolation of RNA using guanidium slats. Inclusion of a reductant enhances denaturation by breaking intramolecular protein disulfide bonds Meth. Enzymol. 152, 219 (1987) 5. Bacarese-Hamilton, A., Prevents the oxidation of cholecystokinin during its extraction from brain and duodenal tissues Ann. N. Y. Acad. Sci. 448, 571 (1985) 6. Jabusch, J.R., In protein sequence analysis Anal. Biochem. 91, 532 (1978) |