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Two‐dimensional Analysis of Skim Milk Proteins Using Preparative Isoelectric Focusing Followed by Polyacrylamide Gel Electrophoresis
| Content Provider | Semantic Scholar |
|---|---|
| Author | Kim, Hye Young Jiménez-Flores, Rafael |
| Copyright Year | 1992 |
| Abstract | A preparative isoelectric focusing (IEF) method was applied to separate skim milk proteins using the Rotofor device in a pH 3–10 gradient containing 4 M urea/1% triton X-100. Each of the 20 fractions obtained from the Rotofor device was then analyzed by polyacrylamide gel electrophoresis (PAGE). Both urea-PAGE and SDS-PAGE were used to separate purified caseins and skim milk resulting in comparable two-dimensional patterns. The major bovine caseins (αs1, αs2, β, K-casein) were resolved better on urea-PAGE. The αs1-and β-casein were focused at pH ∼ 4.5 and 4.8, respectively, whereas αs2-caseins focused as several bands at pH 6.2–6.8. The A variant of K-casein focuses at Fractions 6–9 which is slightly more acidic than the B variant that focuses at Fractions 7–13. No sample pretreatment was necessary to analyze skim milk proteins and urea-PAGE clearly resolved bands of all major caseins and whey proteins. Preparative isoelectric focusing followed by PAGE was found to be a useful and powerful method to analyze milk proteins in two-dimensions. This technique facilitates the analysis of the relative amounts of proteins in milk, as well as simplifies the detection of changes and foreign proteins in milk. |
| Starting Page | 307 |
| Ending Page | 321 |
| Page Count | 15 |
| File Format | PDF HTM / HTML |
| DOI | 10.1111/j.1745-4514.1992.tb00454.x |
| Volume Number | 16 |
| Alternate Webpage(s) | http://digitalcommons.calpoly.edu/cgi/viewcontent.cgi?article=1059&context=dsci_fac |
| Alternate Webpage(s) | https://digitalcommons.calpoly.edu/cgi/viewcontent.cgi?article=1059&context=dsci_fac&httpsredir=1&referer= |
| Alternate Webpage(s) | https://doi.org/10.1111/j.1745-4514.1992.tb00454.x |
| Language | English |
| Access Restriction | Open |
| Content Type | Text |
| Resource Type | Article |
