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A highly sensitive enzyme-linked immunosorbent assay for quantification of adipocytokines secreted by mouse adipocytes

Paper ID Volume ID Publish Year Pages File Format Full-Text
4315 220 2009 6 PDF Available
Title
A highly sensitive enzyme-linked immunosorbent assay for quantification of adipocytokines secreted by mouse adipocytes
Abstract

The mouse preadipocyte 3T3-L1 line is the most useful cell line for the study of adipocytes. Adipocytes secrete adipocytokines, and abnormal adipocytokine production can cause the metabolic syndrome. Although it is important to understand the characteristics of mouse adipocytokine secretion, it is difficult to quantify these products because they are produced in low concentrations. Here, we developed a highly sensitive enzyme-linked immunosorbent assay (ELISA) for detecting the concentrations of mouse adipocytokines, such as TNFα and leptin. In this method, the antigen was sandwiched by using goat- and rabbit-derived polyclonal antibodies, and the fluorescence intensity produced in the reaction with 4-methylumbelliferyl-β-galacto-sidase pyranosidase (MUG) was measured. TNFα and leptin could be measured at concentrations as low as approximately 1 pg/ml. By using our ELISA method, we also measured the concentrations of TNFα and leptin in mouse 3T3-L1 preadipocytes and adipocytes. The differentiation of preadipocytes into adipocytes enhanced TNFα production and secretion and reduced the leptin production. The amount of TNFα produced in the adipocytes was 3.0 ng/mg-protein; this amount was considerably higher than that of leptin.

Keywords
ELISA; TNF; Leptin; Adipocytes
First Page Preview
A highly sensitive enzyme-linked immunosorbent assay for quantification of adipocytokines secreted by mouse adipocytes
Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 43, Issue 1, 15 January 2009, Pages 58–63
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering