The use of quantitative immunocytochemistry (QICC) to measure calbindin D28k-like immunoreactivity in the rat brain.
We have developed a method of quantitative immunocytochemistry using an iodinated second antibody to visualise the anatomical distribution of primary antibodies in tissue sections, by macroautoradiography. Computer-assisted densitometry was used to analyse the pattern of optical densities within autoradiograms. The amount of antigen present in tissue sections was then quantified by comparison with non-biological standards which were processed in parallel with the tissue sections. Using this technique we have measured calbindin like-immunoreactivity in 4 areas of rat brain and have found that the values obtained are similar to those obtained by radioimmunoassay. A similar approach can be used to quantify autoradiograms by comparison with antigen standards to measure amounts of radiolabelled immunoreactivity and determine concentrations of biologically active molecules in discrete brain areas.
Davenport
A P
AP
Department of Medicine, University of Cambridge, Addenbrooke's Hospital, UK.
Augood
S J
SJ
Lawson
D E
DE
Emson
P C
PC
D000818:Animals; D001345:Autoradiography; D001923:Brain Chemistry; D064092:Calbindin 1; D064026:Calbindins; D002645:Chickens; D003720:Densitometry; D007150:Immunohistochemistry; D051381:Rats; D064030:S100 Calcium Binding Protein G / Q000032:analysis*
C576500:Calb1 protein, rat; D064092:Calbindin 1; D064026:Calbindins; D064030:S100 Calcium Binding Protein G