How do I prepare a 1 M DTT solution from powder and how should I store the solution?
Preparing 1 M DTT:
1) Dissolve 3.09 g DTT in 20 mL distilled H2O.
2) Dispense into 1 mL aliquots.
3) Store the aliquots at -20 degrees C.
Note: Upon thawing the vial, there may be some precipitation that can be re-dissolved by warming and mixing.
Introduction of sulfhydryl groups into proteins at carboxyl sites.
Authors:Lin CM, Mihal KA, Krueger RJ
Journal:Biochim Biophys Acta
PubMed ID:2160279
'A two-step procedure for introduction of sulfhydryl groups at protein carboxyl groups is described. The resultant proteins contain 2-aminoethanethiol residues bound by amide linkages to the protein carboxyl groups. First an amide bond is formed between a carboxyl group of the protein and one of the amino groups of cystamine. ... More
Simple alkanethiol groups for temporary blocking of sulfhydryl groups of enzymes.
Authors:Smith DJ, Maggio, ET, Kenyon GL
Journal:Biochemistry
PubMed ID:163643
'New reagents for the temporary blocking of active or accessible sulfhydryl groups of enzymes have been developed. These reagents, which are either alkyl alkanethiolsulfonates or alkoxycarbonylalkyl disulfides, rapidly and quantitatively place various RS- groups on the sulfhydryls to generate mixed disulfides. In all cases native enzymes can be regenerated with ... More
An introduction to methods for analyzing thiols and disulfides: Reactions, reagents, and practical considerations.
Authors:Hansen RE, Winther JR,
Journal:Anal Biochem
PubMed ID:19664585
'This review outlines the basic issues to consider when dealing with biochemical and cellular aspects of thiol–disulfide chemistry. The overall focus is on practical aspects, including typical biochemical experimental conditions and caveats to consider in interpreting results. Reagents for thiol derivatization and disulfide reduction are evaluated and compared, and we ... More
Measurement of cyst(e)amine in physiological samples by high performance liquid chromatography.
Authors:Garcia RA, Hirschberger LL, Stipanuk MH
Journal:Anal Biochem
PubMed ID:3394941
'Two methods for measurement of cyst(e)amine in physiological samples are described. One method involves reduction of disulfides present in the sample with tributylphosphine, reversed phase chromatography of thiols, and electrochemical detection of cysteamine and other thiols. The other method involves reduction of disulfides with dithiothreitol, derivatization of thiols with 7-diethylamino-3-(4''-maleimidylphenyl)-4-methylcoumarin, ... More