Characterization of a monoclonal antibody prepared against plant actin.
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.
Southern Regional Research Center, New Orleans, Louisiana.
J M
JM
Andersland
D D
DD
Fisher
C L
CL
Wymer
R J
RJ
Cyr
M V
MV
Parthasarathy
D008841:Actin Cytoskeleton / Q000737:chemistry* / Q000648:ultrastructure; D000199:Actins / Q000032:analysis / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity; D015638:Cytochalasin D / Q000494:pharmacology; D003368:Gossypium; D006825:Hybridomas; D015151:Immunoblotting; D007150:Immunohistochemistry; D008840:Microfilament Proteins / Q000187:drug effects / Q000648:ultrastructure; D008856:Microscopy, Fluorescence; D018532:Pisum sativum; D010947:Plants, Toxic; D011058:Pollen; D011523:Protoplasts / Q000648:ultrastructure; D014026:Nicotiana / Q000648:ultrastructure; D003313:Zea mays
D000199:Actins; D000911:Antibodies, Monoclonal; D008840:Microfilament Proteins; D015638:Cytochalasin D