Characterization of a monoclonal antibody prepared against plant actin

Cell Motil Cytoskeleton. 1994;29(4):339-44. doi: 10.1002/cm.970290406.

Abstract

Anti-actin monoclonal antibodies were prepared using phalloidin-stabilized actin that was purified from pea roots by DNase I affinity chromatography. One monoclonal antibody, designated mAb3H11, bound plant actin in preliminary screenings and was further analyzed. Immunoblot analysis showed that this antibody had a high affinity for plant actin in crude and purified preparations but a low affinity for rabbit muscle actin. In immunoblots of plant extracts separated on two-dimensional gels it appeared to bind all actin isoforms recognized by the JLA20 anti-chicken actin antibody. Using immunofluorescent cytochemistry, the antibody was used to observe actin filaments in aldehyde-fixed and methanol-treated tobacco protoplasts. These results indicate that mAb3H11 should be a useful reagent for the study of plant actins.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Actin Cytoskeleton / chemistry*
  • Actin Cytoskeleton / ultrastructure
  • Actins / analysis
  • Actins / immunology*
  • Antibodies, Monoclonal / immunology*
  • Antibodies, Monoclonal / isolation & purification*
  • Antibody Specificity
  • Cytochalasin D / pharmacology
  • Gossypium
  • Hybridomas
  • Immunoblotting
  • Immunohistochemistry
  • Microfilament Proteins / drug effects
  • Microfilament Proteins / ultrastructure
  • Microscopy, Fluorescence
  • Peas
  • Plants, Toxic
  • Pollen
  • Protoplasts / ultrastructure
  • Tobacco / ultrastructure
  • Zea mays

Substances

  • Actins
  • Antibodies, Monoclonal
  • Microfilament Proteins
  • Cytochalasin D