Purification and Electrophoretic Characterization of Lactate Dehydrogenase from Mammalian Blood: A Different Twist on a Classic Experiment
| العنوان: | Purification and Electrophoretic Characterization of Lactate Dehydrogenase from Mammalian Blood: A Different Twist on a Classic Experiment |
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| المؤلفون: | Linda S. Brunauer |
| المصدر: | Journal of Chemical Education. 93:1108-1114 |
| بيانات النشر: | American Chemical Society (ACS), 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, chemistry.chemical_classification, Bioanalysis, Chromatography, biology, Chemistry, Elution, education, 05 social sciences, Size-exclusion chromatography, 050301 education, General Chemistry, Enzyme assay, Education, 03 medical and health sciences, chemistry.chemical_compound, 030104 developmental biology, Enzyme, Affinity chromatography, Chemical engineering, Lactate dehydrogenase, Protein purification, biology.protein, 0503 education |
| الوصف: | A multiweek protein purification suite, suitable for upper-division biochemistry or biotechnology undergraduate students, is described. Students work in small teams to isolate the enzyme lactate dehydrogenase (LDH) from a nontraditional tissue source, mammalian blood, using a sequence of three column chromatographic procedures: ion-exchange, size exclusion, and affinity chromatography. Protein and enzyme activity elution profiles are determined by graphical analysis of assay data collected using rapid microplate spectrophotometric assays. Students perform quantitative assays on LDH pools and use these data to build a purification table for use in evaluating the protocol. The protocol typically generates final overall fold-purifications from 1500 to 2500 and activity recoveries of 45–60%. Electrophoretic separations in both denaturing and native-gel format are analyzed both visually and by use of commercial digital analysis software to assess the isoenzyme pattern of isolated LDH and to further evaluate th... |
| تدمد: | 1938-1328 0021-9584 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_________::1054ad53114adf45a65c2494f58ddb09Test https://doi.org/10.1021/acs.jchemed.5b00713Test |
| رقم الانضمام: | edsair.doi...........1054ad53114adf45a65c2494f58ddb09 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 19381328 00219584 |
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