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Introduction to Protein Electrophoresis

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Electrophoretic Separation of Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1855))

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Abstract

This chapter briefly discusses the developments in electrophoresis of proteins from Tiselius’ moving-boundary electrophoresis to the modern-day two-dimensional polyacrylamide gel electrophoresis. It also touches upon the staining methods used to visualize total proteins post electrophoresis.

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References

  1. Tiselius A (1938) Elektrophoretische Messungen am Eiweiss. Kolloid-Ztschr 85:129

    Article  CAS  Google Scholar 

  2. Righetti PG (2005) Electrophoresis: the march of pennies, the march of dimes. J Chromatogr AJ Chromatogr A 1079:24–40

    Article  CAS  Google Scholar 

  3. Von Klobusitzky P, Konig P (1939) Biochemische Studien uber die Gifte der Schlan gengattung. Arch Exp Path Pharm 192:271–275

    Article  CAS  Google Scholar 

  4. Evered DF (1959) Ionophoresis of acidic and basic amino acids on filter paper using low voltages. Biochim Biophys Acta 36:14–19

    Article  CAS  Google Scholar 

  5. Ingram VM (1958) The comparison of normal human and sickle-cell haemoglobins by fingerprinting. Biochim Biophys Acta 28:539–545

    Article  CAS  Google Scholar 

  6. Smithies O (1955) Zone electrophoresis in starch gels: group variations in serum proteins of normal human adults. Biochem J 61:629–641

    Article  CAS  Google Scholar 

  7. Poulik MD (1957) Starch gel electrophoresis in a discontinuous system of buffers. Nature 180:1477–1479

    Article  CAS  Google Scholar 

  8. Raymond S, Weintraub L (1959) Acrylamide gel as a supporting medium for zone electrophoresis. Science 130:711–712

    Article  CAS  Google Scholar 

  9. Raymond S (1964) Acrylamide gel electrophoresis. Ann N Y Acad Sci 121:35–65

    Google Scholar 

  10. Ornstein L (1964) Disc electrophoresis.I. Background and theory. Ann N Y Acad Sci 28:32–49

    Google Scholar 

  11. Gelfi C, Righetti PG (1981) Polymerization kinetics of polyacrylamide gels I. Effect of different cross-linkers. Electrophoresis 2:213–219

    Article  CAS  Google Scholar 

  12. Righetti PG, Gelfi C, Bosisio AB (1981) Polymerization kinetics of polyacrylamide gels III. Effect of catalysis. Electrophoresis 2:291–295

    Article  CAS  Google Scholar 

  13. Andrews AT (1986) Polyacrylamide gel electrophoresis (PAGE):homogenous gel and buffer systems. In: Electrophoresis:theory, techniques, and biochemical and clinical applications. Oxford University Press, New York

    Google Scholar 

  14. Maurer HR (1971) Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis, 2nd edn. Walter de Gruyter, Berlin, New York

    Google Scholar 

  15. Margolis J, Kenrick KG (1967) Polyacrylamide gel-electrophoresis across a molecular sieve gradient. Nature 214:1334–1336

    Article  CAS  Google Scholar 

  16. Margolis J, Kenrick KG (1967) Electrophoresis in polyacrylamide concentration gradient. Biochem Biophys Res Commun 27:68–73

    Article  CAS  Google Scholar 

  17. Shapiro AL, Vinuela E, Maizel JV (1967) Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun 28:815–820

    Article  CAS  Google Scholar 

  18. Ornstein L (1964) Disc electrophoresis. I. Background and theory. Ann N Y Acad Sci 121:321–349

    Article  CAS  Google Scholar 

  19. Davis BJ (1964) Disc electrophoresis. II. Method and application to human serum proteins. Ann N Y Acad Sci 121:404–427

    Article  CAS  Google Scholar 

  20. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  21. Steinberg TH (2009) Protein gel staining methods: an introduction and overview. Methods Enzymol 463:541–563

    Article  CAS  Google Scholar 

  22. Neuhoff V, Stamm R, Hansjorg E (1985) Clear background and highly sensitive protein staining with Coomassie blue dyes in polyacrylamide gels: a systematic analysis. Electrophoresis 6:427–488

    Article  CAS  Google Scholar 

  23. Switzer RC, Merril CR, Shifrin S (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal Biochem 98:231–237

    Article  CAS  Google Scholar 

  24. Daban J-R, Bartolome S, Samso M (1991) Use of the hydrophobic probe Nile red for the fluorescent staining of protein bands in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem 199:169–174

    Article  CAS  Google Scholar 

  25. Berggren K, Chernokalskaya E, Steinberg TH et al (2000) Background-free, high sensitivity staining of proteins in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gels using a luminescent ruthenium complex. Electrophoresis 21:2509–2521

    Article  CAS  Google Scholar 

  26. Mackintosh JA, Choi H-Y, Bae S-H et al (2003) A fluorescent natural product for ultrasensitive detection of proteins in one-dimensional and two-dimensional gel electrophoresis. Proteomics 3:2273–2288

    Article  CAS  Google Scholar 

  27. Steinberg TH, Agnew BJ, Gee KR et al (2003) Global quantitative phosphoprotein analysis using multiplexed proteomics technology. Proteomics 3:1128–1144

    Article  CAS  Google Scholar 

  28. Kinoshita E, Kinoshita-Kikuta E, Takiyama K, Koike T (2006) Phosphate binding tag, a new tool to visualize phosphorylated proteins. Mol Cell Proteomics 5:749–757

    Article  CAS  Google Scholar 

  29. O’Farrell PH (1975) High resolution two-dimensional electrophoresis of proteins. J Biol Chem 250:4007–4021

    PubMed  PubMed Central  Google Scholar 

  30. Jorgenson JW, Lukacs KD (1981) Free-zone electrophoresis in glass capillaries. Anal Chem 53:1298–1302

    Article  CAS  Google Scholar 

  31. Sisu E, Flangea C, Serb A, Rizzi A, Zamfir AD (2011) High-performance separation techniques hyphenated to mass spectrometry for ganglioside analysis. Electrophoresis 32:1591–1609

    CAS  PubMed  Google Scholar 

  32. Stepanova S, Kasicka V (2016) Recent applications of capillary electromigration methods to separation and analysis of proteins. Anal Chim Acta 933:23–42

    Article  CAS  Google Scholar 

  33. Zhu Y, Li Z, Wang P, Shen L, Zhang D, Yamaguchi Y (2018) Factors affecting the separation performance of proteins in capillary electrophoresis. J Chromatogr B 1083:63–67

    Article  CAS  Google Scholar 

  34. Gilges M, Kleemiss MH, Schomburg G (1994) Capillary zone electrophoresis separations of basic and acidic proteins using poly(vinyl alcohol) coatings in fused silica capillaries. Anal Chem 66:2038–2046

    Article  CAS  Google Scholar 

  35. Novotny MV, Cobb KA, Liu JP (1990) Recent advances in capillary electrophoresis of proteins, peptides and amino acids. Electrophoresis 11:735–749

    Article  CAS  Google Scholar 

  36. Huhn C, Ramautar R, Wuhrer M, Somsen GW (2010) Relevance and use of capillary coatings in capillary electrophoresis-mass spectrometry. Anal Bioanal Chem 396:297–314

    Article  CAS  Google Scholar 

  37. Horvath J, Dolnik V (2001) Polymer wall coatings for capillary electrophoresis. Electrophoresis 22:644–655

    Article  CAS  Google Scholar 

  38. Dawod LM, Arvin NE, Kennedy RT (2017) Recent advances in protein analysis by capillary and microchip electrophoresis. Analyst 142:1847–1866

    Article  CAS  Google Scholar 

  39. Breadmore MC, Shallan AI, Rabanes HR, Gstoettenmayr D, Keyon AS, Gaspar A, Dawod M, Quirino JP (2013) Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2010–2012). Electrophoresis 34:29–54

    Article  CAS  Google Scholar 

  40. Breadmore MC, Thabano JRE, Dawod M, Kazarian AA, Quirino JP, Guijt RM (2009) Recent advances in enhancing the sensitivity of electrophoresis and electrochromatography in capillaries and microchips (2006–2008). Electrophoresis 30:230–248

    Article  CAS  Google Scholar 

  41. Dawod M, Chung DS (2011) High-sensitivity capillary and microchip electrophoresis using electrokinetic supercharging. J Sep Sci 34:2790–2799

    Article  CAS  Google Scholar 

  42. Polikarpov N, Potolytsyna V, Bessonova E, Tripp S, Appelhans D, Voit B, Kartsova L (2015) Dendritic glycopolymers as dynamic and covalent coating in capillary electrophoresis: view on protein separation processes and detection of nanogram-scaled albumin in biological samples. J Chromatogr A 1378:65–73

    Article  CAS  Google Scholar 

  43. Zhu Z, Lu JJ, Liu S (2012) Protein separation by capillary gel electrophoresis: a review. Anal Chim Acta 709:21–31

    Article  CAS  Google Scholar 

  44. Wu D, Regnier FE (1992) Sodium dodecyl sulfate-capillary gel electrophoresis of proteins using non-cross-linked polyacrylamide. J Chromatogr 608:349–356

    Article  CAS  Google Scholar 

  45. Ouimet CM, Shao H, Rauch JN, Dawod M, Nordhues B, Dickey CA, Gestwicki JE, Kennedy RT (2016) Protein cross-linking capillary electrophoresis (PXCE) for protein-protein interaction analysis. Anal Chem 88:8272–8278

    Article  CAS  Google Scholar 

  46. Agostino FJ, Krylov SN (2015) Advances in steady-state continuous-flow purification by small-scale free-flow electrophoresis. TrAC Trends Anal Chem 72:68–79

    Article  CAS  Google Scholar 

  47. Krivankova L, Boceck P (1998) Continuous free-flow electrophoresis. Electrophoresis 19:1064–1074

    Article  CAS  Google Scholar 

  48. Hannig K (1978) Continuous free-flow electrophoresis as an analytical and preparative method in biology. J Chromatogr 1978(159):183–191

    Article  Google Scholar 

  49. Hannig K (1982) New aspects in preparative and analytical continuous and free-flow electrophoresis (a review). Electrophoresis 3:235–243

    Article  CAS  Google Scholar 

  50. Shen QY, Guo CJ, Yan J, Zhang Q, Xie HY, Jahan S, Fan LY, Xiao H, Cao CX (2015) Target protein separation and preparation by free-flow electrophoresiscoupledwith charge-to-mass ratio analysis. J Chromatogr A 1397:73–80

    Article  CAS  Google Scholar 

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Correspondence to Pothur R. Srinivas .

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Srinivas, P.R. (2019). Introduction to Protein Electrophoresis. In: Kurien, B., Scofield, R. (eds) Electrophoretic Separation of Proteins. Methods in Molecular Biology, vol 1855. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8793-1_2

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  • DOI: https://doi.org/10.1007/978-1-4939-8793-1_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8792-4

  • Online ISBN: 978-1-4939-8793-1

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