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A donor of thiol groups used in biochemical and pharmacologic studies.
Synonym(s): Cleland reagent


W. Wallace, U.S. biochemist, 1930–.
Cleland reagent - a substance used to reduce disulfide bonds in proteins. Synonym(s): dithiothreitol
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5,1 mM EDTA, 1 M NaCl, 20% glycerol) containing 2 mM P-mercaptoethanol, and then against buffer B containing 1 mM dithiothreitol.
Protein bands in the appropriate relative molecular weight range were manually excised, reduced with 5 mM dithiothreitol (DTT) and alkylated with 50 mM iodoacetamide (Bio-Rad Laboratories), digested with trypsin (Promega Corp.
A20-[micro]L aliquot was diluted to 300 [micro]L with 50 mmol/L ammonium bicarbonate (pH 8), reduced with 10 mmol/L dithiothreitol (DTT) at 60[degrees]C for 45 min, alkylated with 10 mmol/L iodoacetamide at room temperature for 30 min in the dark, reduced again with 5 mmol/L DTT at room temperature for 30 min in the dark, and digested overnight at 37[degrees]C with modified trypsin at a 1:50 (wt/wt) trypsin:total protein ratio.
2) and lysed in a lysis buffer (8 M urea, 4% CHAPS, 40 mM dithiothreitol and 0.
0, 1 mM phenylmethylsulfonyl fluoride (PMSF) (Roche, Germany), 50 x Protease inhibitor (Roche, Germany), 200 mM dithiothreitol (DTT) (GE Healthcare, Sweden).
6 [micro]L of 1 tool dithiothreitol at 65[degrees]C for 15 min, neutralized with 8.
4]), dithiothreitol (DTT), propyl thiouracil (PTU) were purchased from Sigma Chemical Co.
3]), trifluoroethanol (TFE), iodoacetamide (IAA), trifluoroacetic acid (TFA), and TPCK-treated trypsin (T-1426) from Sigma-Aldrich; dithiothreitol (DTT) and formic acid from Fluka; fetal calf serum (FCS) from Invitrogen (Gibco 10437-028); Zwittergent detergent 3-16 (Z 3-16) from Calbiochem; and highly purified A1AT reference material from Athens Research & Technology (16-16-011609).
4], 1 mM dithiothreitol, the desired concentration of extract, and an NADPH-generating system composed of 0.
The beads were resuspended in 100 [micro]L of 10 mmol/L dithiothreitol and incubated at 56 [degrees]C for 45 min.