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Journal of Lipid Research, Vol 34, 325-330, Copyright © 1993 by Lipid Research, Inc.
ZF Stephan and EC Yurachek
DiI-LDL (3,3'-dioctadecylindocarbocyanine-low density lipoprotein) has been
extensively used in morphological and microscopic studies of
receptor-mediated metabolism of LDL in many cell lines. To date the use of
this fluorescent probe in a quantitative assay of LDL receptor activity has
not been widely used in studies with multiple samples due to the lack of a
practical method for quantitatively recovering cell- associated DiI.
Therefore, detection by 125I-labeled LDL has remained the method of choice
for assaying LDL receptor activity rapidly and reliably. In this paper, we
describe a rapid, simple, and nonradioactive assay of LDL receptor activity
using DiI-LDL. The increased sensitivity of this method was achieved by
modifications to the labeling procedure of LDL and to the extraction of DiI
from cells for subsequent fluorescence determination. These modifications
did not affect the affinity of DiI-LDL toward HepG2 cells, and the assay
was easily adapted to a rapid screen for LDL receptor modulators in this
cell model.
ARTICLES
Rapid fluorometric assay of LDL receptor activity by DiI-labeled LDL
Research Department, CIBA-GEIGY Corporation, Summit, NJ 07901.
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