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

Link to article in PubMed

PubMed ID

7130194