P. Venembre et al., COMPARISON OF ENHANCED CHEMILUMINESCENCE AND COLORIMETRIC TECHNIQUES FOR THE IMMUNO-DETECTION OF ALPHA(1)-ANTITRYPSIN, Clinica chimica acta, 227(1-2), 1994, pp. 175-184
In order to detect, characterize and quantify blotted proteins, such a
s human alpha(1)-antitrypsin (AAT), there is a need for a specific, ex
tremely sensitive, non-radioactive and uniform revelation system appli
cable to diluted biological fluids and to culture supernatants of cell
s isolated from such fluids. We compared two immunochemical revelation
systems, enhanced chemiluminescence (ECL) and colorimetric procedures
, applied to human AAT, after determining their optimal conditions of
performance. ECL was the most sensitive method (down to 50 pg blotted
AAT), but could not be used to quantify AAT. In contrast, the colorime
tric method enables quantification of blotted AAT, either simply dotte
d or transferred after SDS-polyacrylamide gel electrophoresis, but is
not as sensitive as ECL. Using these two complementary procedures, we
have been able to detect AAT in the culture supernatant of a monocytic
cell line (THP-1), to characterize the different forms of AAT present
in the culture supernatant of blood monocytes and to quantify both.