References on casein electrophoresis
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- Snezana Jovanovic, Miroljub Barac, Ognjen Macej, Tanja Vucic, and Caslav Lacnjevac. SDS-PAGE Analysis of Soluble Proteins in Reconstituted Milk Exposed to Different Heat Treatments. Sensors (Basel). 2007 Mar; 7(3): 371–383.
- "The soluble protein composition of the treated skim milk samples was detected by the SDS-polyacrilamide gel electrophoresis (SDS-PAGE) performed according to Fling and Gregerson [16] on 12.5 % and 15% slab gel. Prior to the electrophoresis, soluble proteins have been diluted to 2 mg/cm3 with the sample buffer (pH 6, 8). Two types of diluted samples have been prepared, with and without 2-mercaptoethanol (0.05%).2-Mercaptoethanol was used as dissociating reagent to obtain degradation of co-aggregates possibly formed by disulfide interactions."
- "most of the soluble co-aggregates are the result of the disulfide interactions. Namely, after the use of 2-mercaptoethanol, the most intensive bands of co-aggregates (141 kDa and 71 kDa-bands, Fig.3, Fig.4.1) have disappeared almost completely."
- Marcelo F. Pardo and Claudia L. Natalucci. Electrophoretic Analysis (Tricine-SDS-PAGE) of Bovine Caseins. Acta Farm. Bonaerense 21 (1): 57-60 (2002)
- "Traditionally Urea-PAGE methods have been preferred for the study of milk proteins, owed to this dissociating agent prevents molecular aggregation. Notwithstanding, Urea-PAGE gels afford a poor resolution (specially the α-caseins), the bands are rounded and often diffuse, specially in the αs2-casein region. Moreover, κ- casein is not detectable (Figure 1a).
- On the contrary, in the Tricine-SDS-PAGE method here reported the resolution was considerably improved with respect to Urea-PAGE methods, as well as SDS-PAGE and gradient PAGE methods. In the new method, αs2-, αs1-, β- and κ-casein are visualized as sharp bands (Figure 1c)."
- "Casein solution was prepared by dissolving 1g casein in 100 mL of 100 mM Tris-HCl buffer (pH 8.0) and heating in a boiling bath for 20 min. The solution was filtrated without cooling and stored at 4 °C until to be used. Casein solutions must be discarded after 2 days.
- Casein solutions were diluted by adding an equal volume of double-concentrated sample buffer (125 mM Tris-HCl, pH 6.8, 4% SDS, 10% 2-mercaptoethanol, 0,4% bromophenol blue and 20% glycerol) and heated at 100 °C for 5 min, centrifuged and immediately stored at -20 °C until analysis. Low molecular weight standards (Bio-Rad) were prepared in sample buffer. Samples and standards (2.5-5 µl) were applied under the cathodic buffer using a 10 µl Hamilton micrometer syringe."