Purification of undegraded ceruloplasmin from outdated human plasma
| Autor: | Mathys M.J. Oosthuizen, Leonie Nel, Robert L. Crookes, Johannes A. Myburgh |
|---|---|
| Rok vydání: | 1985 |
| Předmět: |
Time Factors
Chemical Phenomena Biophysics Biochemistry Specimen Handling Humans Centrifugation Amino Acids Equilibrium Centrifugation Molecular Biology Polyacrylamide gel electrophoresis Gel electrophoresis Chromatography biology Isoelectric focusing Elution Chemistry Ceruloplasmin Cell Biology Isoelectric point biology.protein Electrophoresis Polyacrylamide Gel Isoelectric Focusing Ultracentrifugation |
| Zdroj: | Analytical Biochemistry. 146:1-6 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/0003-2697(85)90386-0 |
| Popis: | A method for the rapid isolation of homogeneous undegraded ceruloplasmin from outdated human plasma is reported. The procedure consists of a precipitation step with polyethylene glycol 4000, batchwise adsorption and elution from QAE-Sephadex, and gradient elution from DEAE-Sepharose CL-6B. Ceruloplasmin was purified 1740-fold and the yield from outdated plasma was 67%. The purified ceruloplasmin was found to be homogeneous on anionic polyacrylamide gel electrophoresis (PAGE), sodium dodecyl sulfate-PAGE, isoelectric focusing, and low-speed equilibrium centrifugation. The isoelectric point as determined by isoelectric focusing was 4.4. The purified enzyme was sensitive to storage; when a sample was resubmitted to PAGE after 4 months of storage at 4°C, two bands were obtained and the fast-moving band showed no oxidase activity. The molecular weight estimated by gel electrophoresis and sedimentation equilibrium centrifugation was 130,000. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|