Characterization of a monoclonal antibody prepared against plant actin

✍ Scribed by Andersland, John M. ;Fisher, Deborah D. ;Wymer, Carol L. ;Cyr, Richard J. ;Parthasarathy, M. V.

• Publisher: John Wiley and Sons
• Year: 1994
• Tongue: English
• Weight: 537 KB
• Volume: 29
• Category: Article
• ISSN: 0886-1544
• DOI: 10.1002/cm.970290406

✦ Synopsis

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 mAb3H 1 1, 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 JLAZO 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. o 1994 Wiley-Lisa, Inc.