Protein Blotting of Basic Proteins Resolved on Acid-Urea-Triton-Polyacrylamide Gels

  • Geneviève P. Delcuve
  • James R. Davie
Part of the Springer Protocols Handbooks book series (SPH)


The electrophoretic resolution of histones on acetic acid-urea-Triton (AUT) polyacrylamide gels is the method of choice to separate basic proteins, such as histone variants, modified histone species, and high-mobility group proteins 14 and 17 (1-6 and see  Chapters 16 and  17). Basic proteins are resolved in this system on the basis of their size, charge, and hydrophobicity. In previous studies, we analyzed the abundance of ubiquitinated histones by resolving the histones on two-dimensional (AUT into SDS) polyacrylamide gels, followed by their transfer to nitrocellulose membranes, and immunochemical staining of nitrocellulose membranes with an antiubiquitin antibody (7-9). However, transfer of the basic proteins directly from the AUT polyacrylamide gel circumvents the need to run the second-dimension SDS gel and accomplishes the analysis of several histone samples. We have described a method that efficiently transfers basic proteins from AUT polyacrylamide gels to nitrocellulose membranes (10). This method has been used in the immunochemical detection of modified histone, isoforms, and histone H1 subtypes (6,11-13).


Nitrocellulose Membrane Basic Protein Equilibration Buffer Histone Variant Transfer Buffer 
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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Geneviève P. Delcuve
    • 1
  • James R. Davie
    • 1
  1. 1.Manitoba Institute of Cell BiologyWinnipegCanada

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