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Most immunofluorescence methods rely on techniques dealing with a very large number of cells. However, when the number of cells in a sample is low (e.g., when cumulus cells must be analyzed from individual cumulus-oocyte complexes), specific techniques are required to conserve, fix, and analyze cells individually. We established and validated a simple and effective method for collecting and immobilizing low numbers of cumulus cells that enables easy and quick quantitative immunofluorescence analysis of proteins from individual cells. To illustrate this technique, we stained proprotein of a disintegrin and metalloproteinase with thrombospondin-like repeats-1 (proADAMTS-1) and analyzed its levels in individual porcine cumulus cells.

Original publication

DOI

10.1016/j.ab.2015.04.003

Type

Journal article

Journal

Anal Biochem

Publication Date

01/07/2015

Volume

480

Pages

31 - 33

Keywords

Cell-Tak, Cumulus cells, Immobilization, Protein immunostaining, proADAMTS-1, Animals, Cumulus Cells, Disintegrins, Fluorescent Antibody Technique, Matrix Metalloproteinases, Swine