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Immunogold labelling is a quantitative method as demonstrated by studies on aminopeptidase N in microvillar membrane
G H Hansen1, L L Wetterberg, H Sjöström
1Department of Biochemistry C, Panum Institute, University of Copenhagen, Denmark.
Abstract:
Microvillar membrane vesicle preparations with varying content of aminopeptidase N were prepared from enterocytes of the pig small intestine. Postembedding immunogold labelling of aminopeptidase N was performed on these glutaraldehyde/paraformaldehyde-fixed, osmium tetroxide-treated and Epon-embedded microvillar membrane vesicles. The number of gold particles per micrometre microvillar membrane (labelling intensity) was calculated and compared to the corresponding enzymatic activity. A very close relationship was found between labelling intensity and aminopeptidase N activity, demonstrating that postembedding immunogold labelling can be used quantitatively.