Skip to main content
SpringerLink
Log in

Fluorescent Staining of Gels

  • Protocol
  • First Online:
Protein Electrophoresis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 869))

Abstract

Certain transition metal complexes show intensive fluorescence when bound to proteins. They can be used to stain gels after electrophoresis with a sensitivity approaching that of silver staining, but in a much simpler and more reproducible procedure. Stains can be prepared easily and at a fraction of the cost of commercially available reagents.

Hydrophobic dyes can be used to stain gels without fixing; they do not interfere with later blotting or electro-elution.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover 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

References

  1. Ornstein L (1964) Disc electrophoresis – I. Background and theory. Ann N Y Acad Sci 121:321–349, http://www.pipeline.com/∼lenornst/DiscEle1.pdf

    Article  PubMed  CAS  Google Scholar 

  2. Davis BJ (1964) Disc electrophoresis – II. Method and application to human serum protein. Ann N Y Acad Sci 121:404–427, http://www.pipeline.com/∼lenornst/DiscEle2.pdf

    Article  PubMed  CAS  Google Scholar 

  3. Fazekas de St. Groth S, Webster RG, Datyner A (1963) Two new staining procedures for quantitative estimation of proteins on electrophoretic strips. Biochim Biophys Acta 71:377–391. doi:10.1016/0006-3002(63)91092-8

    Article  Google Scholar 

  4. Heukeshoven J, Dernick R (1988) Improved silver staining procedure for fast staining in phastsystem development unit. I. staining of sodium dodecyl sulfate gels. Electrophoresis 9:28–32. doi:10.1002/elps.1150090106

    Article  PubMed  CAS  Google Scholar 

  5. Merril CR, Goldman D, Sedman SA, Ebert MH (1981) Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211:1437–1438. doi:10.1126/science.6162199

    Article  PubMed  CAS  Google Scholar 

  6. Buxbaum E (2010) Biophysical chemistry of proteins: an introduction to laboratory methods. Springer, New York. ISBN ISBN 978-1-4419-7250-7

    Google Scholar 

  7. Daban J-R, Bartolomé S, Samsó 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. doi:10.1016/0003-2697(91)90085-8

    Article  PubMed  CAS  Google Scholar 

  8. Miller I, Crawford J, Gianazza E (2006) Protein stains for proteomics application: which, when, why? Electrophoresis 6:5385–5408. doi:10.1002/pmic.200600323

    CAS  Google Scholar 

  9. Rabilloud T, Strub J-M, Luche S, Dorsselaer AV, Lunardi J (2001) A comparison between Sypro ruby and ruthenium II tris(bathophenantroline disulfonate) as fluorescent stains for protein detection in gels. Proteomics 1:699–704. doi:10.1002/elps.1150130190

    Article  PubMed  CAS  Google Scholar 

  10. Pluder F, Beck-Sickinger AG (2007) One-step procedure for staining of proteins with ruthenium II tris(bathophenanthroline disulfonate). Anal Biochem 361:299–301. doi:10.1016/j.ab.2006.11.003

    Article  PubMed  CAS  Google Scholar 

  11. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, http://bioinfcpcri.org/misc/nature.pdf

    Article  PubMed  CAS  Google Scholar 

  12. Alba FJ, Bermudez A, Bartolome S, Daban J-R (1996) Detection of five nanograms of protein by two-minute nile red staining of unfixed SDS gels. Biotechniques 21:625–626, http://www.biotechniques.com/multimedia/archive/ 00009/96214bm12_9719a.pdf

    PubMed  CAS  Google Scholar 

  13. Smith I, Cromie R, Stainsby K (1988) Seeing gel wells well. Anal Biochem 169:370–371. doi:10.1016/0003-2697(88)90297-7

    Article  PubMed  CAS  Google Scholar 

  14. Cleland WW (1964) Dithiothreitol, a new protective reagent for SH groups. Biochemistry 3:480–482. doi:10.1021/bi00892a002

    Article  PubMed  CAS  Google Scholar 

  15. Crans DC, Zhang B, Gaidamauskas E, Keramidas AD, Willsky GR, Roberts CR (2010) Is vanadate reduced by thiols under biological conditions? changing the redox potential of V(V)/V(IV) by complexation in aqueous solution. Inorg Chem 49:4245–4256. doi:10.1021/ic100080k

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by funding from Ross University.

Author information

Authors and Affiliations

  1. Ross University School of Medicine, Roseau, Commonwealth of Dominica, West Indies

    Engelbert Buxbaum

Authors
  1. Engelbert Buxbaum
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Engelbert Buxbaum .

Editor information

Editors and Affiliations

  1. Oklahoma Medical Research Foundation, Univ. of Oklahoma Health Sciences Center, NE. 13th Street 825, Oklahoma City, 73104, Oklahoma, USA

    Biji T. Kurien

  2. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, Oklahoma, USA

    R. Hal Scofield

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Buxbaum, E. (2012). Fluorescent Staining of Gels. In: Kurien, B., Scofield, R. (eds) Protein Electrophoresis. Methods in Molecular Biology, vol 869. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-821-4_48

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-821-4_48

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-820-7

  • Online ISBN: 978-1-61779-821-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation