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Quantitative Measurement of GLUT4 Translocation to the Plasma Membrane by Flow Cytometry
Published on: November 7, 2010
Quantitation of GLUT1 and GLUT4 mRNA using a solution hybridization assay
D A Meier1, D Pastorek, R G James
1Department of Medicine, Medical College of Wisconsin, Milwaukee 53226.
Abstract:
The development of a solution hybridization assay for detecting GLUT1 and GLUT4 mRNA is described. The details of this assay are described in which copy RNA is used to quantitate messenger RNA in total RNA samples. This solution hybridization assay is highly specific and reproducible and is significantly more sensitive than Northern blotting. Since GLUT mRNAs can be quantitated in as little as 25 mg tissue, this technique is essential when the supply of tissue is limited. Furthermore, the elimination of gel-based separation techniques allows for mRNA quantitation in several hundred samples within two days following isolation of samples.
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