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Protein Electrophoresis

  • Paul Richards
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

There are an estimated 100,000 genes in the entire human genome, 10% of which may be expressed in a particular cell-type at a given time Purlfying and analyzing the products of genes (proteins), from humans and other organisms, offers a unique challenge to the blochemlst and one that is essential to our understanding of blologlcal function The diversity in size, lomc charge, solublhty, stability, cellular expression, and posttranslatlonal modlficatlon of proteins requires protein-analysis techniques that have high-resolving capacities Electrophoretlc separation techniques provide a hlghresolution, adaptable and simple range of methodologies used in nearly all forms of protein analysis (1)

Keywords

Native Page Silver Cation Entire Human Genome Carrier Ampholyte Trichloroacetic Acid Solution 
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.

References

  1. 1.
    Dunn, M. J. (1993) Gel Elecfrophoresis: Proteins. Bios, Oxford, UK.Google Scholar
  2. 2.
    Estep, T. N. and Miller, T. J. (1986) Optimisation of erythrocyte membrane glycoprotein fluorescent labelling with dansylhydrazine after polyacrylarnide gel electrophoresis. Anal.Biochem. 157, 100–105.PubMedCrossRefGoogle Scholar
  3. 3.
    Debruyne, I. (1983) Staining of alkali-labile phophoproteins and alkaline phosphates on polyacrylamide gels. Anal. Biochem. 133, 110–115.PubMedCrossRefGoogle Scholar
  4. 4.
    Tsai, C.M. and Frasch, C. E. (1982) A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal. Biochem. 119, 115–119.PubMedCrossRefGoogle Scholar
  5. 5.
    Bag-Hansen, T C (1993) Immunoelectrophoresls, in Gel Electrophoreszs of Protezns (Hames, B D and Rickwood, D, eds), IRL, Oxford, UK, pp 273–300Google Scholar
  6. 6.
    Raymond, S and Wemtraud, L S (1959) Acrylamlde gel as a supporting medmm for zone electrophoresls Sczence 130,711–721CrossRefGoogle Scholar
  7. 7.
    O’Connel, P B H and Brady, C J (1976) Polyacrylamlde gels with modified crosslinkages Anal Blochem 76,63–73CrossRefGoogle Scholar
  8. 8.
    Rodbard, D, Chrambach, A, and Wels, G H (1974) in Electrophorests and Zsoelectrzc Focuszng zn Polyacrylamzde Gel (Allen, R C and Mauvrer, H R, eds), Walter de Grugtner, Berlm, p 62Google Scholar
  9. 9.
    Heussen, C and Dowdle, E B (1980) Electrophorectlc analysis of plasmmogen actlvtators in polyacrylamlde gel containing sodium dodecyl sulphate and copolymerlsed substrates Anal Bzoche 102, 196–202CrossRefGoogle Scholar
  10. 10.
    Gabriel, O (1971) Locating enzymes on gels Methods Enzymol 22,578–604CrossRefGoogle Scholar
  11. 11.
    Dunn, M J (1993) Electrophoresls under native condltlons, in Gel Electrophoreszs Protezns, BIOS, Oxford, UK, pp 31–39Google Scholar
  12. 12.
    Carr-Eusbe, D, Lederer, F, DlCp, K H, and Elsevler, S in (1991) Processing of the precursor of protamme P2 in mouse Blochem J 277,39–45Google Scholar
  13. 13.
    Rlghettl, P G (1983) Isoelectric Focuszng Theory Methodology and Applzcatzons Elsevler, AmsterdamGoogle Scholar
  14. 14.
    Weber, K and Osborn, M (1969) The rellablhty of molecular weight determmatlon by dodecyl sulphate polyacrylamlde gel electrophoresls J BtoZ Chem 244,4406–4412Google Scholar
  15. 15.
    Laemmh, U, K (1970) Cleavage of structural proteins during the assembly of the head of the bacterlopahge T4 Nature 227,680–685CrossRefGoogle Scholar
  16. 16.
    Schagger, H and von Jagow, G (1987) Tncme-sodium dodecyl sulfate-polyacrylamlde gel electrophoresls for the separation of proteins in the range from 1 to 100 KDa Anal Blochem 166,368–379CrossRefGoogle Scholar
  17. 17.
    Poduslo, J F and Rodbard, D (1982) Molecular weight estimation using sodmm dodecyl sulphate-pore gradient electrophoresls Anal Blochem 101,394–406CrossRefGoogle Scholar
  18. 18.
    Wilson, C in (1983) Staming of proteins on gels comparisons of dyes and procedures Methods Enzymol 91,236–246PubMedCrossRefGoogle Scholar
  19. 19.
    Neuhoff, V, Arold, N, Taube, D, and Ehrhardt, W (1988) Improved staming of proteins in polyacrylamlde gels mcluding lsoelectrlc focusing gels with clear background and nanogram sensltlvlty using Coomassle brllllant blue G-250 and R-250 Electrophoreszs 9,255–262CrossRefGoogle Scholar
  20. 20.
    Ward, A H and Allen, W S (1972) An improved procedure for protein staming in polyacrylamlde gels with a new type of Coomassle bnlhant blue Anal Bzochem 48,617–620CrossRefGoogle Scholar
  21. 21.
    Oakley, B R, Klrsch, D R, and Morris, N R (1980) A slmphfied ultrasensItIve sliver stain for detecting protems in polyacrylamlde gels Anal Bzochem 105,361–363CrossRefGoogle Scholar
  22. 22.
    Bonner, W in and Laskey, R A (1974) A film detectton method for tntmm-labelled protems and nucleic actds in polyacrylamlde gels Eur J Blochem 46, 83–88Google Scholar
  23. 23.
    Cleveland, D W, Fischer, S G, Klrschner, M W, and Laemmh, U K (1977) Peptlde mapping by limited proteolysls in sodium dodecyl sulfate and analysis by gel electro-1 phoresls J Bzol Chem 252,1102–1106Google Scholar
  24. 24.
    Gultekm, H and Heermann, K H (1988) The use of polyvmyhdenedlfluorlde membranes as a general blotting matrix Anal Bzochem 172, 320–329CrossRefGoogle Scholar
  25. 25.
    Kennedy, T E, Wager-Smith, K, Barzllal, A., Kandel, E R, and Sweatt, J D (1988)Sequencing proteins from acrylamme gels Nature 336,499,500PubMedCrossRefGoogle Scholar
  26. 26.
    Pluskal, M G, Przekop, M B, Kavoman, M R, Vecoh, C, and Hicks, D A (1986)ImmobllonTM PVDF transfer membrane a new membrane substrate for Western blotting of proteins BloTechntques 4,272–283Google Scholar
  27. 27.
    Hochstrasser, D F, Harrington, M G, Hochstrasser, A-C, Miller, M J, and Menl, C R (1988) Methods for increasing the resolution of two-dlmenslonal protem electrophore-SISAnal Blochem 173,424–435CrossRefGoogle Scholar
  28. 28.
    Perdew, G H, Scaup, H, and Sehvonchlck, D P (1983) The use of detergents in twodimensIona gel electrophoresls of trout-lrver mlcrosomes Anal Bzochem 135,453–455CrossRefGoogle Scholar
  29. 29.
    Gorg, A (1993) Two-dImensiona electrophoresls with lmmoblhzed pH gradients current state Bzochem Sot Trans 21, 130–133Google Scholar

Copyright information

© Humana Press Inc , Totowa, NJ. 1998

Authors and Affiliations

  • Paul Richards
    • 1
  1. 1.Leicester UnzversztyUK

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