Words similar to mercaptoethanol
Example sentences for: mercaptoethanol
How can you use “mercaptoethanol” in a sentence? Here are some example sentences to help you improve your vocabulary:
1 mM 2-mercaptoethanol/1% penicillin-streptomycin) at 37°C/5% CO 2 until analysis.
Each construct was transformed into E. coli strain BL21 (DE3) (Novagen), and protein was expressed, purified by glutathione-agarose affinity chromatography, cleaved from GST and stored in 20 mM Tris (pH 7.6 at 4°C), 50% glycerol, 1 mM EGTA, 0.1% 2-mercaptoethanol, 4 mM MnCl 2 at -20°C as previously described for PP5 [ 18 ] . Recombinant Ppt1p and Ppt1p (1-504) each contain an additional seven residues, Gly-Ser-Gly-Ser-Glu-Phe-Asp, which are remnants of the thrombin-cleavage site.
The serine/threonine protein phosphatase type 5 (PP5) was purified from bovine brain according to the following procedure: a bovine brain was homogenized in ~ 3 volumes of buffer A (50 mM HEPES pH 7.5 4°C, 1 mM EDTA, 10 mM PMSF and 0.1% 2-mercaptoethanol) and subjected to centrifugation at 15,000 g for 40 min. followed by a 45%-60% ammonium sulfate fractionation of the supernatant.
The eluate from the SP column was then diluted 5-fold with buffer C (20 mM tris pH 8.5, 1 mM EDTA and 0.1% 2-mercaptoethanol) and subjected to anion-exchange chromatography on a BioScale Q column (2 ml from BioRad).
32P-labeled substrates were prepared and assays performed as described previously [ 18 ] . Briefly, protein phosphatase reactions contained 2 or 4 nM enzyme in a final volume of 30 μl containing buffer A (50 mM Tris, pH 7.6, 1 mM EDTA, 1 mM EGTA, 0.1% 2-mercaptoethanol), 0.4% ethanol and 10 μM radiolabeled phosphosubstrate in the absence or presence of AA [ 19 ] . Reactions were initiated by adding substrate and lipid, and incubated at 30°C for 10 to 15 min.
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