Skip to main content
SpringerLink
Log in

Gelelektrophoresen

  • Chapter
Gentechnische Methoden

  • 20k Accesses

Zusammenfassung

Die Wanderung geladener Teilchen im elektrischen Feld wird zur Trennung komplexer Gemische von Biomolekülen ausgenutzt. Die Trennmethoden werden unter dem Begriff Elektrophorese zusammengefasst. Da die meisten Biomoleküle Ladungen tragen, gehören Elektrophoresen zu den wichtigsten analytischen Methoden. Für die Untersuchung und Charakterisierung komplexer Mischungen sowie zur Überprüfung ihrer Einheitlichkeit sind sie unentbehrlich. Die wesentlichen Unterschiede in den Trennungsmethoden ergeben sich aus dem für jedes Biomolekül charakteristischem Verhältnis von Ladung zu Masse bzw. aus ihrer Molekülgröße und -form.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 54.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 69.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Similar content being viewed by others

Literatur

  • Cooper, T.G. (1981): Biochemische Arbeitsmethoden. Walter de Gruyter, Berlin.

    Google Scholar 

  • Lottspeich, F., Zorbas, H. (Hrsg.) (1998): Bioanalytik. Spektrum Akademischer Verlag, Heidelberg, Berlin.

    Google Scholar 

  • Süßmuth, R., Eberspächer, J., Haag, R., Springer, W. (1999): Mikrobiologisch-Biochemisches Praktikum. Georg Thieme Verlag, Stuttgart.

    Google Scholar 

Literatur

  • Coligan, J.E., Dunn, B.M., Ploegh, H.L., Speicher, D.W., Wingfield, P.T. (Hrsg.) (1995, 1996): Current Protocols in Protein Science. John Wiley & Sons, New York.

    Google Scholar 

  • Hames, B.D., Rickwood, D. (Hrsg.) (1990): Gel Electrophoresis of Proteins: A Practical Approach. IRL Press at OUP, Oxford.

    Google Scholar 

  • Kellner, R., Lottspeich, F., Meyer, H.E. (Hrsg.) (1999): Microcharacterization of Proteins. VCH, Weinheim.

    Google Scholar 

Literatur

  • Sambrook, J., Fritsch, E.F., Maniatis, T. (1989): Molecular Cloning-A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

  • Rigby, P.W., Dieckmann, M., Rhodes, C., Berg, P. (1977): Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA-polymerase I. J. Mol. Biol. 113(1), 237–251.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Feinberg, A.P., Vogelstein, B. (1984): A Technique for Radiolabeling DNA Restriction Endonuclease Fragments to High Specific Activity. Anal. Biochem. 137, 266–267.

    Article  PubMed  CAS  Google Scholar 

  • Keller, G.H. (1989): DNA Probes. Stockten Press, New York.

    Google Scholar 

  • The DIG System User’s Guide for Filter Hybridization (1995): Boehringer Mannheim GmbH Biochemica.

    Google Scholar 

Literatur

  • Süßmuth, R., Eberspächer, J., Haag, R., Springer, W. (1999): Mikrobiologisch-Biochemisches Praktikum. Georg Thieme Verlag, Stuttgart.

    Google Scholar 

  • Sambrook, J., Fritsch, E.F., Maniatis, T. (1989): Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press.

    Google Scholar 

Literatur

  • Sambrook, J., Fritsch, E.F., Maniatis, T. (1989): Molecular Cloning. A Laboratory Manual. Cold Spring Harbor Laboratory Press.

    Google Scholar 

Literatur

  • Mary K.B. Berlyn (1998): Linkage Map of Escherichia coli K-12, Edition 10: The Traditional Map. Mikrobiology and Molecular Biology Reviews, 62, 814–984.

    Google Scholar 

Literatur

  • Bast, E. (2001): Mikrobiologische Methoden. 2. Aufl. Spektrum Akademischer Verlag, Heidelberg.

    Google Scholar 

Literatur

  • Coligan J., Dunn B.M., Ploegh H.L., Speicher D.W., Wingfield P.T. (Hrsg.) (1995): Current Protocols in Protein Science Volume 1. John Wiley & Sons, New York, Kap. 6, 8, 9.

    Google Scholar 

  • Deutscher M.S. (1990): Principles in Protein Purification. Methods Enzymol. 182.

    Google Scholar 

  • Scopes, R.K. (1987): Protein Purification, Principles and Practice. 2nd ed. Springer Verlag, New York.

    Google Scholar 

Literatur

  • Gallagher, S.R. (1995) in: Coligan, J.E., Dunn, B.M., Ploegh, H.L., Speicher, D.W., Wingfield, P.T. (Hrsg.): One-Dimensional SDS Gel Electrophoresis of Proteins. Current Protocols in Protein Science. John Wiley & Sons, New York.

    Google Scholar 

  • Laemmli, U.K. (1970): Cleavage of Structural Proteins During the Assembly of the Head of Bacteriophage T4. Nature 227, 680–685.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Gallagher, S.R. (1995) in: Coligan, J.E., Dunn, B.M., Ploegh, H.L., Speicher, D.W., Wingfield, P.T. (Hrsg.): One-Dimensional SDS Gel Electrophoresis of Proteins. Current Protocols in Protein Science. John Wiley & Sons, New York.

    Google Scholar 

  • Schägger, H., von Jagow, G. (1987): Tricine-Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis for the Separation of Proteins in the Range from 1 to 100 kDa. Anal. Biochem. 166, 368–379.

    Article  PubMed  Google Scholar 

Literatur

  • Weber, K., Pringle, J.R., Osborn, M. (1972): Measurement of Molecular Weights by Electrophoresis on SDSAcrylamide Gel. Methods Enzymol. 26, 3–27.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Rothe, G.M., Purkhanbaba, H. (1982): Determination of Molecular Weights and Stokes’ Radii of Non-Denatured Proteins by Polyacrylamide Gradient Gel Electrophoresis 1. An Equation Relating Total Polymer Concentration, the Molecular Weight of Proteins in the Range of 104–106, and Duration of Electrophoresis. Electrophoresis 3, 33–42.

    Article  CAS  Google Scholar 

  • Walker, J. M. (2002): Gradient SDS Polyacrylamide Gel Electrophoresis of Proteins in: Walker, J. M. (Hrsg.) The Protein Protocols Handbook. Humana Press Inc., Totowa, NJ, 69–72.

    Chapter  Google Scholar 

Literatur

  • Akin, D.T., Shapira, R., Kinkade, J.M.Jr. (1985): The Determination of Molecular Weights of Biologically Active Proteins by Cetyltrimethylammonium Bromide Polyacrylamide Gel Electrophoresis. Anal. Biochem. 145, 170–176.

    Article  PubMed  CAS  Google Scholar 

  • Andrews, A.T. (1986): Electrophoresis. Theory, Techniques and Biochemical and Clinical Applications. 2nd ed. Oxford University Press, New York.

    Google Scholar 

  • Eley, M.H., Burns, P.C., Kannapell, C.C., Campbell, P.S. (1979): Cetyltrimethylammonium Bromide Polyacrylamide Gel Electrophoresis: Estimation of Protein Subunit Molecular Weights Using Cationic Detergents. Anal. Biochem. 92, 411–419.

    Article  PubMed  CAS  Google Scholar 

  • Schägger, H., Cramer, W.A., von Jagow, G. (1994): Analysis of Molecular Masses and Oligomeric States of Protein Complexes by Blue Native Electrophoresis and Isolation of Membrane Protein Complexes by Two-Dimensional Native Electrophoresis. Anal. Biochem. 217, 220–230.

    Article  PubMed  Google Scholar 

  • Schägger, H., von Jagow, G. (1991): Blue Native Electrophoresis for Isolation of Membrane Protein Complexes in Enzymatically Active form. Anal. Biochem. 199, 223–231.

    Article  PubMed  Google Scholar 

Literatur

  • Patel, D., Rickwood, D. (1996) in: Price, N.C. (Hrsg.): Proteins Labfax. BIOS Scientific Publishers Ltd, Oxford.

    Google Scholar 

  • Radola, B.J. (1983) in: Tschesche, H. (Hrsg.): Modern Methods in Protein Chemistry. Walter de Gruyter, Berlin, 21–48.

    Google Scholar 

  • Strahler, J.R., Hanash, S.M., Somerlot, L., Weser, J., Postel, W., Görg, A. (1987): High Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis of Basic Myeloid Polypeptides: Use of Immobilized pH Gradients in the First Dimension. Electrophoresis 8, 165–173.

    Article  CAS  Google Scholar 

  • Strahler, J.R., Hanash, S.M. (1991): Immobilized pH Gradients. Analytical and Preparative Use. Methods 3, 109–114.

    Article  CAS  Google Scholar 

Literatur

  • Dunbar, B.S. (1987) in: Dunbar, B.S. (Hrsg.): Two-Dimensional Electrophoresis and Immunological Techniques. Plenum, New York, 173–195.

    Chapter  Google Scholar 

  • Dunn, M.J., Burghes, A.H.M. (1983): High Resolution Two-Dimensional Polyacrylamide Gel Electrophoresis I. Methodological Procedures. Electrophoresis 4, 97–116.

    Article  CAS  Google Scholar 

  • Gianazza, E. (2002): Casting Immobilized pH Gradients (IPGs) in: Walker, J. M. (Hrsg.) The Protein Protocols Handbook. Humana Press Inc., Totowa, NJ, 169–180.

    Chapter  Google Scholar 

  • Gravel, P. (2002): Two-Dimensional Polyacrylamide Gel Electrophoresis of proteins using Carrier Ampholyte pH Gradients in the First Dimension in: Walker, J.M. (Hrsg.) The Protein Protocols Handbook. Humana Press Inc., Totowa, NJ, 163–168.

    Chapter  Google Scholar 

  • Hochstrasser, D.F., Harrington, M.C., Hochstrasser, A.C., Miller, M.J., Merril, C.R. (1988): Methods for Increasing the Resolution of Two-Dimensional Protein Electrophoresis. Anal. Biochem. 173, 424–435.

    Article  PubMed  CAS  Google Scholar 

  • Klose, J. (1975): Humangenetik 26, 231–243.

    PubMed  CAS  Google Scholar 

  • Lottspeich, F., Zorbas, H. (Hrsg.) (1998): Bioanalytik. Spektrum Akademischer Verlag, Heidelberg, Berlin.

    Google Scholar 

  • O’Farrell, P.H. (1975): High Resolution Two-Dimensional Electrophoresis of Proteins. J. Biol. Chem. 250, 4007–4021.

    CAS  Google Scholar 

  • Young, D.A., Voris, B.P., Maytin, E.V., Colbert, R.A. (1983): Very-High-Resolution Two-Dimensional Electrophoretic Separation of Proteins on Giant Gels. Methods Enzymol. 91, 190–214.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Pharmacia LKB Sonderdruck (1988): Arbeitsanleitung. Herstellung von Gelen für das PhastSystem.

    Google Scholar 

Literatur

  • Rabilloud, T. (1992): A Comparison between Low Background Silver Diamine and Silver Nitrate Protein Stains. Electrophoresis 13, 429–439.

    Article  PubMed  CAS  Google Scholar 

  • Wilson, C. (1983): Staining of Proteins on Gels. Comparisons of Dyes and Procedures. Methods Enzymol. 91, 236–247.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Ansorge, W. (1985): Fast and Sensitive Detection of Protein and DNA Bands by Treatment with Potassium Permanganate. J. Biochem. Biophys. Meth. 11, 13–20.

    Article  PubMed  CAS  Google Scholar 

  • Heukeshoven, J., Dernick, R. (1985): Simplified Method for Silver Staining of Proteins in Polyacrylamide Gels and the Mechanism of Silver Staining. Electrophoresis 6, 103–112.

    Article  CAS  Google Scholar 

  • Merril, C.R., Dunau, M.L., Goldman, D. (1981): A Rapid Sensitive Silver Stain for Polypeptides in Polyacrylamide Gels. Anal. Biochem. 110, 201–207.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Alwine, J.C. (1979): Detection of Specific RNAs or Specific Fragments of DNA by Fractionation in Gels and Transfer to Diazobenzyloxymethyl Paper. Methods Enzymol. 68, 220–242.

    Article  PubMed  CAS  Google Scholar 

  • Lunn, G., Sansone, E. (1987): Ethidium bromide. Destruction and Decontamination of Solutions. Anal. Biochem. 162, 453–458.

    Article  PubMed  CAS  Google Scholar 

  • SYBR® Green I Nucleic Acid Gel Stain, Product Information Sheet, MP 7567 07/16/96, Molecular Probes (Invitrogen GmbH).

    Google Scholar 

  • SYBR® Safe DNA Gel Stain, Product Information, MP 33100 06/19/2007, Molecular Probes (Invitrogen GmbH)

    Google Scholar 

  • GelRed™ Nucleic Acid Gel Stain, Product Information, Jan 23, 2009, Biotium (Biotrend Chemikalien GmbH).

    Google Scholar 

  • GelGreen™ Nucleic Acid Gel Stain, Product Information, Apr 9, 2010, Biotium (Biotrend Chemikalien GmbH).

    Google Scholar 

Literatur

  • Blin, N., Gabain, A.V., Bujard, H. (1975): Isolation of Large Molecular Weight DNA from Agarose Gels for Further Digestion by Restriction Enzymes. FEBS Lett. 53, 84–86.

    Article  PubMed  CAS  Google Scholar 

  • Chen, Z., Ruffner, D.E. (1996): Modified Crush-and-Soak Method for Recovering Oligodeoxynucleotides from Polyacrylamide Gel. Biotechniques 21, 820–822.

    PubMed  CAS  Google Scholar 

  • Moore, D., Chory, M., Ribaudo, R.K. (1994). In: Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (Hrsg.): Isolation and Purification of Large DNA Restriction Fragments from Agarose Gels. Current Protocols in Molecular Biology. John Wiley & Sons, New York.

    Google Scholar 

  • Thuring, R.W., Sanders, J.B., Borst, P.A. (1975): A Freeze-Squeeze Method for Recovering Long DNA from Agarose Gels. Anal. Biochem. 66, 213–220.

    Article  PubMed  CAS  Google Scholar 

  • Vogelstein, B., Gillespie, D. (1979): Preparative and Analytical Purification of DNA from Agarose. Proc. Natl. Acad. Sci. USA. 76, 615–619.

    Article  PubMed  CAS  Google Scholar 

  • Wienand, U., Schwarz, Z., Felix, G. (1978): Electrophoretic Elution of Nucleic Acids from Gels Adapted for Subsequent Biological Tests. Application for Analysis of mRNAs from Maize Endosperm. FEBS Lett. 98, 319–323.

    Article  Google Scholar 

Literatur

  • SYPRO Protein Gel Stains, Molecular Probes (Invitrogen GmbH) MP 6650 11/27/95.

    Google Scholar 

Literatur

  • Eng, P.R., Parkes, C.O. (1974): SDS Electrophoresis of Fluorescamine-Labeled Proteins. Anal. Biochem. 59, 323–325.

    Article  PubMed  CAS  Google Scholar 

  • Ragland, W.L., Pace, J.L., Kemper, D.L. (1974): Fluorometric Scanning of Fluorescamine-Labeled Proteins in Polyacrylamide Gels. Anal. Biochem. 59, 24–33.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • Higgins, R.C., Dahmus, M.E. (1979): Rapid Visualization of Protein Bands in Preparative SDS-Polyacrylamide Gels. Anal. Biochem. 93, 257–260.

    Article  PubMed  CAS  Google Scholar 

  • Jenö, P., Horst, M. (2002): Electroelution of Proteins from Polyacrylamide Gels in: Walker, J. M. (Hrsg.) The Protein Protocols Handbook, Humana Press Inc., Totowa, NJ, 299–305.

    Chapter  Google Scholar 

Literatur

  • Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (Hrsg.) (1998): Current Protocols in Molecular Biology. John Wiley & Sons, New York.

    Google Scholar 

  • Rickwood, D., Hames, B.D. (Hrsg.) (1990): Gel Electrophoresis of Nucleic Acids — A Practical Approach. IRL Press, Washington D.C.

    Google Scholar 

  • Sambrook, J., Russel, D.W. (2001): Molecular Cloning — A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

Literatur

  • Birren, B.W., Lai, E., Clark, S.M., Hood, L., Simon, M.I. (1988): Optimized Conditions for Pulsed Field Gel Electrophoretic Separations of DNA. Nucl. Acids Res. 16, 7563–7582.

    Article  PubMed  CAS  Google Scholar 

  • Carle, G.F., Frank, M., Olson, M.V. (1986): Electrophoretic Separations of Large DNA Molecules by Periodic Inversion of the Electric Field. Science 232, 65–68.

    Article  PubMed  CAS  Google Scholar 

  • Finney, M. (1998): Pulsed-Field-Gel Electrophoresis in: Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (Hrsg.) Current Protocols in Molecular Biology. Green Publishing Associates and Wiley Interscience, John Wiley & Sons, New York, Kap. 2.5.16.

    Google Scholar 

  • Schwartz, D.C., Cantor, C.R. (1984): Separation of Yeast Chromosome-Sized DNAs by Pulsed Field Gradient Gel Electrophoresis. Cell 37, 67–75.

    Article  PubMed  CAS  Google Scholar 

Literatur

  • McDonell, M.W., Simon, M.N., Studier, F.W. (1977): Analysis of Restriction Fragments of T7 DNA and Determination of Molecular Weights by Electrophoresis in Neutral and Alkaline Gels. J. Mol. Biol. 110, 119–146.

    Article  PubMed  CAS  Google Scholar 

  • Odgen, R.C., Adams, D.A. (1987): Electrophoresis in Agarose and Acrylamide Gels. Methods Enzymol. 152, 61–87.

    Article  Google Scholar 

Literatur

  • Goldberg, D.A. (1980): Isolation and Partial Characterization of the Drosophila Alcohol Dehydrogenase Gene. Proc. Natl. Acad. Sci. USA 77, 5794–5798.

    Article  PubMed  CAS  Google Scholar 

  • Liu, Y.-C., Chou, Y.-C. (1990): Formaldehyde in Formaldehyde/Agarose Gel May be Eliminated without Affecting the Electrophoretic Separation of RNA Molecules. Biotechniques 9 (5), 558.

    PubMed  CAS  Google Scholar 

  • McMaster, G.K., Carmichael, G.G. (1977): Analysis of Single-and Doublestranded Nucleic Acids on Polyacrylamide and Agarose Gels by Using Glyoxal and Acridine Orange. Proc. Natl. Acad. Sci. USA 74, 4835–4838.

    Article  PubMed  CAS  Google Scholar 

  • Rickwood, D., Hames, B.D. (Hrsg.) (1990): Gel Electrophoresis of Nucleic Acids — A Practical Approach. IRL Press, Washington D.C.

    Google Scholar 

Literatur

  • Ellington, A., Pollard, J.D.Jr. (1998) in: Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G., Smith, J.A., Struhl, K. (Hrsg.): Purification of Oligonucleotides Using Denaturing Polyacrylamide Gel Electrophoresis. Current Protocols in Molecular Biology. John Wiley & Sons, New York.

    Google Scholar 

  • Peacock, A.C., Digman, C.W. (1968): Molecular Weight Estimation and Separation of Ribonucleic Acid by Electrophoresis in Agarose-Acrylamide Composite Gels. Biochemistry 7, 668–674.

    Article  PubMed  CAS  Google Scholar 

  • Rickwood, D., Hames, B.D. (Hrsg.) (1990): Gel Electrophoresis of Nucleic Acids — A Practical Approach. IRL Press, Washington D.C.

    Google Scholar 

  • Sambrook, J., Russel, D.W. (2001): Molecular Cloning — A Laboratory Manual. 3rd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York.

    Google Scholar 

Download references

Authors
  1. Ute Dechert
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Monika Jansohn Sophie Rothhämel

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Spektrum Akademischer Verlag Heidelberg

About this chapter

Cite this chapter

Dechert, U. (2012). Gelelektrophoresen. In: Jansohn, M., Rothhämel, S. (eds) Gentechnische Methoden. Spektrum Akademischer Verlag, Heidelberg. https://doi.org/10.1007/978-3-8274-2430-3_2

Download citation

Publish with us

Policies and ethics

Search

Navigation