Concentrated Tris solutions for the preparation, depolymerization, and assay of actin: application to erythroid actin.
Anal Biochem
; 225(2): 291-5, 1995 Mar 01.
Article
en En
| MEDLINE
| ID: mdl-7762794
ABSTRACT
High concentrations of Tris are effective in dissociating actin-containing complexes, such as the red cell membrane cytoskeleton. A preparative procedure for red cell actin is based on the dissociation of the membrane skeletal complex in a buffer containing 1 M Tris hydrochloride, followed by gel filtration chromatography in the same medium. The actin is recovered as the monomer and is fully native, as judged by its critical concentration of polymerization, inhibition of DNase I, stimulation of myosin ATPase, and the appearance in the electron microscope of filaments, both bare and decorated with heavy meromyosin, and of magnesium ion-induced paracrystals. The Tris solution causes rapid depolymerization of F-actin with no denaturation, and the solution of monomeric actin in this medium is stable for many weeks in the cold; concentrated Tris is more reliable than guanidinium chloride for the depolymerization of F-actin in the estimation of total actin concentration by the DNase I inhibition assay.
Buscar en Google
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Trometamina
/
Actinas
/
Eritrocitos
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Anal Biochem
Año:
1995
Tipo del documento:
Article
País de afiliación:
Reino Unido