A Trichloroacetic Acid–Acetone Method Greatly Reduces Infrared Autofluorescence of Protein Extracts from Plant Tissue

  • Shultz R
  • Settlage S
  • Hanley-bowdoin L
 et al. 
  • 21

    Readers

    Mendeley users who have this article in their library.
  • 17

    Citations

    Citations of this article.

Abstract

Immunoblotting is used to determine many important characteristics of proteins. After electrophoretic separation, proteins are transferred to a membrane and reacted with a specific antibody. The antibody-protein complex is then visualized by radiographic, chromogenic, chemiluminescent, or more recently described fluorescence detection meth- ods. Fluorescence-based detection offers some advantages over other approaches, includ- ing increased sensitivity, improved quantifiable range, and the ability to detect multiple antigens on the same blot. However, this technique is unavailable for analysis of green plant tissues by standard extraction methods because of contaminating autofluorescent pigments. We have compared 3 methods for extracting protein from plant tissue for use with infrared fluorescence-based immunoblot analysis. We report a trichloroacetic acid– acetone method that effectively eliminates autofluorescence while retaining the immuno- genicity of a target protein.

Author-supplied keywords

  • 1
  • abbreviations
  • arabidopsis
  • autofluorescence
  • immunoblot
  • infrared
  • odyssey
  • plant
  • tbst
  • tca
  • trichloroacetic acid
  • tris-buffered saline with 0
  • tween

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • R.W. Shultz

  • S.B. Settlage

  • L. Hanley-bowdoin

  • W.F. Thompson

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free