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."
  • Kang YC, Richardson T. Molecular cloning and expression of bovine kappa-casein in Escherichia coli. J Dairy Sci. 1988 Jan;71(1):29-40.
    "The cell pellet from 1 ml of the overnight culture derived from colonies containing expression plasmids were collected and resuspended in 100 ul of SDS sample buffer. They were boiled for 5 min and 50-ul aliquots for each sample were subjected to 12% SDSPAGE (16). The proteins were blotted on nitrocellulose sheets and tested for the presence of K-casein as described by Hawkes (13) with antibodies raised against K-casein."
  • Hansson L, Bergström S, Hernell O, Lönnerdal B, Nilsson AK, Strömqvist M. Expression of human milk beta-casein in Escherichia coli: comparison of recombinant protein with native isoforms. Protein Expr Purif. 1993 Oct;4(5):373-81.
  • Chong DK, Roberts W, Arakawa T, Illes K, Bagi G, Slattery CW, Langridge WH. Expression of the human milk protein beta-casein in transgenic potato plants. Transgenic Res. 1997 Jul;6(4):289-96.
  • Docena GH, Fernandez R, Chirdo FG, Fossati CA. Identification of casein as the major allergenic and antigenic protein of cow's milk. Allergy. 1996 Jun;51(6):412-6.
    "SDS-PAGE was performed with a Mini-Protean I1 (Bio-Rad) according to Laemmli (13), in 14% polyacrylamide gels. An amount of 35 pg of protein were used per lane, and the run was carried out at 100 V for the stacking gel and at 150 V for 1 h for the running gel."
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