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Analysis of Casein using Two-Dimensional Electrophoresis, Western Blot, and Computer Imaging

  • Marcia F. Goldfarb
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 501)

Abstract

Casein from milk of 20 mothers, 8 weeks postpartum, was analyzed using 2dimensional electrophoresis. The casein micelle under 2-D denaturing conditions gives approximately 15 major spots. I3-Casein, x-casein, para-x-casein, and casein peptides can be tentatively identified by molecular size, isoelectric point, and spot shape characteristics. All spots are not present in all samples. Two spots thought to represent different phosphorylated forms of 13-casein were selected for area and density measurements by means of the NIH-Image program. The larger area at approximately 26 kDa, isoelectric point 5.3, consisted of 1 to 3 closely linked spots, perhaps reflecting phenotype. The smaller spot 24kDa, isoelectric point 5.4, was always a single unit. Visual scanning of gels suggests a wide range of casein concentration. Computer comparison will allow a statistical analysis.

Keywords

Isoelectric Point Human Milk Casein Micelle Major Spot Human Milk Sample 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Anderson L. Two-dimensional Electrophoresis: Operation of the Iso-Dalt System. Washington DC: Large Scale Biology Press; 1991. p. 200.Google Scholar
  2. Goldfarb MF, Savadove MS. Two-dimensional electrophoretic analysis of human milk proteins. Electrophoresis 1989;10:67–70.PubMedCrossRefGoogle Scholar
  3. Goldfarb M. Two-dimensional electrophoretic analysis of human milk-fat-globule membrane proteins with attention to apolipoprotein E patterns. Electrophoresis 1997;18:511–515.CrossRefGoogle Scholar
  4. Kunz C, Lonnerdal B. Casein micelles and casein subunits in human milk. In: Atkinson S, Lonnerdal B, editors. Protein and Non-Protein Nitrogen in Human Milk. Boca Raton: CRC Press; 1989. pp. 10–27.Google Scholar
  5. Lonnerdal B, Atkinson S. Nitrogenous components of milk A. Human milk proteins. In: Jensen RG, editor. Handbook of Milk Composition. San Diego: Academic Press; 1995. pp. 351–494.CrossRefGoogle Scholar
  6. Morrissey JH. Silver stain for proteins in polyacrylamide gels: A modified procedure with enhanced uniform sensitivity. Anal Biochem 1981;17:307–310.Google Scholar
  7. Russ JC. The Image Processing Handbook. 2nded. Boca Raton: CRC Press; 1995. p. 674.Google Scholar
  8. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 1979;76:4350–4354.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2001

Authors and Affiliations

  • Marcia F. Goldfarb
    • 1
  1. 1.Anatek-EPPortland

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