Re-evaluation of Sepharose-insulin as a tool for the study of insulin action. | Zendy

Helmut J. Kolb | Zendy; R. Renner | Zendy; K. Hepp | Zendy; L. Weiß | Zendy; O. Wieland | Zendy

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Re-evaluation of Sepharose-insulin as a tool for the study of insulin action.

Author(s) -

Helmut J. Kolb,

R. Renner,

K. Hepp,

L. Weiß,

O. Wieland

Publication year - 1975

Publication title -

proceedings of the national academy of sciences

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 5.011

H-Index - 771

eISSN - 1091-6490

pISSN - 0027-8424

DOI - 10.1073/pnas.72.1.248

Subject(s) - insulin , sepharose , lipogenesis , biological activity , chemistry , centrifugation , pyruvate dehydrogenase complex , incubation , biochemistry , specific activity , in vitro , endocrinology , biology , adipose tissue , enzyme

The biological activity of Sepharose-insulin in different assays in vitro, e.g., stimulation of glucose oxidation, lipogenesis, and antilipolysis and activation of pyruvate dehydrogenase (EC 1.2.4.1) activity, has been investigated. According to amino acid analysis, between 270 and 330 mug (6.9-8.2 U) of insulin were coupled per ml of packed beads. Related to the total insulin content, 0.2-0.7% of the insulin was biologically active. Comparable biological activity was observed with isolated fat cells and fat pad pieces. After incubation with tissue or cells, Sepharose-insulin particles were separated by centrifugation from the medium. The clear supernatant was assayed for biologically and immunologically reactive insulin and contained soluble insulin activity. A quantitative evaluation of the soluble biological and immunological insulin activity in the supernatant accounted for the total insulin activity of Sepharose-insulin.

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