Optimization of a method for the immunocytochemical detection of nestin in paraffin sections of the rat brain.
The aim of the present work was to develop an optimal and simple protocol for the immunocytochemical detection of nestin (an intermediate filament protein often used as a marker for neural stem cells) in paraffin sections of the rat brain without using an antigen demasking procedure, as this distorts tissue structure. The studies addressed the effects of fixation, the nature of blocking of non-specific antibody binding, the temperature regime, and the incubation time on the results of the immunocytochemical reaction. The results were used to propose a processing protocol allowing clear detection of nestin in rat brain cells with optimal preservation of nervous tissue structure.
Korzhevskii
D E
DE
Department of Morphology, Institute of Experimental Medicine, Russian Academy of Medical Sciences, St. Petersburg.
Gilyarov
A V
AV
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D001923:Brain Chemistry / Q000502:physiology*; D007150:Immunohistochemistry / Q000379:methods*; D007381:Intermediate Filament Proteins / Q000032:analysis*; D019579:Neocortex / Q000033:anatomy & histology / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000032:analysis*; D064231:Nestin; D016612:Paraffin Embedding; D051381:Rats; D013696:Temperature; D016707:Tissue Fixation
D000906:Antibodies; D007381:Intermediate Filament Proteins; D009419:Nerve Tissue Proteins; C577486:Nes protein, rat; D064231:Nestin