Title
A stable, high capacity, F-actin affinity column
UMMS Affiliation
Department of Cell Biology
Date
11-10-1982
Document Type
Article
Subjects
*Actins; Animals; Chromatography, Affinity; Microscopy, Electron; Muscle Proteins; Muscles; Rabbits; Sepharose; Spectrometry, Fluorescence
Disciplines
Cell Biology | Life Sciences | Medicine and Health Sciences
Abstract
A high capacity F-actin affinity matrix is constructed by binding fluorescyl-actin to rabbit anti-fluorescein IgG that is covalently bound to Sepharose 4B. When stabilized with phalloidin, the actin remains associated with the Sepharose beads during repeated washes, activates the ATPase activity of myosin subfragment 1, and specifically binds 125I-heavy meromyosin and 125I-tropomyosin. The associations between the F-actin affinity matrix and the iodinated F-actin binding proteins are monitored both by affinity chromatography and by a rapid, low speed sedimentation assay. Anti-fluorescein IgG-Sepharose should be generally useful as a matrix for the immobilization of proteins containing accessible, covalently bound fluorescein groups.
Rights and Permissions
Citation: J Biol Chem. 1982 Nov 10;257(21):13095-100.
Related Resources
PubMed ID
7130194
