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An isocratic reverse-phase high-performance liquid chromatography method for the separation and quantitation of total pyridine dinucleotides in hepatocyte cultures is described. Cells are extracted with cold 3 M perchloric acid or 0.5 N sodium hydroxide containing 50% (v/v) ethanol and 35% cesium chloride for the determination of the oxidized or reduced pyridine dinucleotides, respectively. Pyridine dinucleotides in the neutralized extracts were separated on an Excellopak ODS C18 (4.6 X 150 mm) column with 0.1 M potassium phosphate, pH 6.0, containing 3.75% methanol as the mobile phase. NAD+ and NADP+ were detected spectrophotometrically at 254 nm. The response was linear from 5 to 4000 pmol with recoveries of NAD+ and NADP+ of 98 and 101.1%, respectively. NADH and NADPH were monitored fluorometrically by activation at 370 nm and emission in the 400-700 nm range. The reduced pyridine dinucleotides had a linear response from 7.5 to 60 pmol with recoveries of NADH and NADPH of 99.4 and 101.3%, respectively. The coefficients of variation for all of the pyridine dinucleotide standards were less than 3.5%. 相似文献
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The lengths of the body, the posterior spine and the three longest anterior spines were measured for 25 specimens of Kellicottia longispina from each of the eight lakes distributed from Imikpuk at Point Barrow, Alaska (latitude 71° 15) to Lake Washington (latitude 47° 38). Collections were available for more than two dates from six of the lakes. Temperature ranged from 1.2° to 18 °C. Mean lengths and ratios were examined in relation to latitude and temperature. Each population differed from the others in some aspect of absolute size, variability, or shape as expressed by the ratios of the dimensions. The population from Point Barrow is similar but not identical to Olofsson's var. heterospina. 相似文献
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Seven polymorphic loci mapping to human chromosomal region 11q22-qter 总被引:11,自引:0,他引:11
Seven polymorphic loci that map to human chromosomal region 11q22-qter are revealed by DNA probes isolated from a chromosome-specific phage library constructed from a human X mouse somatic cell hybrid that has retained an 11q;16q translocation as the only human DNA. Three probes, each of which reveals a two-allele polymorphism, and four probes, each of which detects two linked RFLPs, have been characterized. Using a somatic cell hybrid mapping panel that divides 11q into four discrete sections, the seven clones have been localized to specific chromosomal regions. Localization of one of the clones has been confirmed and refined by in situ hybridization. 相似文献
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