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Title: A novel immunohistochemical semiquantitative technique for endothelial constitutive nitric oxide synthase immunoreactivity in rat coronary artery.
Austin Authors: Zulli, Anthony;Liu, J J
Affiliation: Vascular Biology Unit, Department of Cardiac Surgery, University of Melbourne Austin Hospital, Heidelberg, Australia
Issue Date: 1-Feb-1998
Publication information: The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society; 46(2): 257-62
Abstract: It has been difficult to quantify protein production in small pathological specimens by conventional techniques. We describe a new method for semiquantification of immunohistochemical staining, which involves application of the enzyme-labeled avidin (LAB) technique, coupled with an ultra-sensitive and fast chemiluminescent substrate for alkaline phosphatase. The entire procedure can be completed in less than 3 hr. The final step involves X-ray film exposure for 30 min, and the optical density of the subsequent images is examined with a microcomputer imaging device. The optical densities are translated into relative protein concentrations by a reference standard curve, obtained via an immunoblot. To establish a model for semiquantification of endothelial constitutive nitric oxide synthase (eNOS) protein, we compared the coronary arteries of WKY rats fed a normal chow diet to the coronary arteries of WKY rats fed a cholesterol diet. Using this technique, we have found a relative 130-fold decrease in eNOS in the cholesterol-fed group compared to the normal chow-fed group.
Gov't Doc #: 9446833
Journal: The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society
Type: Journal Article
Subjects: Animals
Cholesterol, Dietary.administration & dosage
Coronary Vessels.enzymology
Endothelium, Vascular.enzymology
Image Processing, Computer-Assisted
Luminescent Measurements
Nitric Oxide Synthase.analysis
Rats, Inbred WKY
Reference Standards
Appears in Collections:Journal articles

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