Words similar to dtt

Example sentences for: dtt

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The resulting protein pellet was resuspended in 10 ml of buffer A (50 mM Tris-HCl, pH 7.5, 5 mM MgCl 2, 0.1 mM EDTA, 0.5 mM DTT, 10 % glycerol and 1 mM PMSF) and dialyzed against 500 ml of buffer A overnight.
SDS-PAGE sample buffer containing 10 mM DTT was added to the washed beads, and the samples were heated at 65°C for 1 min ( 3H fatty acid labeled samples) or boiled for 5 min ( 35S methionine/cysteine labeled samples).
Binding reactions were carried out at room temperature for 30 min in a total volume of 15 μl and contained 2-5 μg of nuclear extracts, 5 μl of 5X gel shift binding buffer (20% glycerol, 5 mM MgCl 2 , 2.5 mM EDTA, 2.5 mM DTT, 250 mM NaCl, 50 mM Tris-HCl, pH 7.5), 2 μg poly (dI-dC) and 3 × 10 4cpm/μl of [ 32P] labeled CREB (5'-AGA GAT TGC CTG ACG TCA GAG AGC TAG-3'), NF-κB (5'-AGT TGA GGG GAC TTT CCC AGG C-3') (Promega Gel Shift Assay Systems, Madison, WI, USA) or CCAAT/enhancer binding protein (C/EBP) (5'-TGC AGA TTG CGC AAT CTG CA-3') (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) oligonucleotides.
An equal mass of each protein sample was subjected to reducing conditions (99°C for 5 min in 100 mM dithiothreitol (DTT)), separated on a 15% SDS-PAGE, and transferred to nitrocellulose membrane according to standard protocols.
All the buffers contained 1 mM DTT.