Determination of Cholesterol content in Cells

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Guy Sovak
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Determination of Cholesterol content in Cells

Dear All,
Here is a nice and easy method for the determination of Cholesterol content in cells.
the protocol is taken from:

Molecular Biology of the Cell
Vol. 15, 31323145, July 2004
Disturbed Cholesterol Traffic but Normal Proteolytic
Function in LAMP-1/LAMP-2 Double-deficient
Fibroblasts?D
Eeva-Liisa Eskelinen,* Christine Katrin Schmidt,* Silja Neu,*
Marion Willenborg,* Graciela Fuertes, Natalia Salvador,
Yoshitaka Tanaka,§ Renate Lu¨ llmann-Rauch, Dieter Hartmann,
Jo¨ rg Heeren,# Kurt von Figura,@ Erwin Knecht, and Paul Saftig* **

The amount of cholesterol was measured in cell extracts prepared using 50
mM Tris, pH 8.0, 2 mM CaCl2, 80 mM NaCl, 1% Triton X-100, with the
Amplex Red Cholesterol Assay kit (Molecular Probes). For the fluorometric
quantification, 25 l of cell extracts and cholesterol standards were incubated
in 96-well plates according to the instructions of the manufacturer, and
fluorescence was detected using Fluorolite1000 (Dynatech Laboratories, Alexandria,
VA).

If any one have got any Question please let me know.
Guy

baobaorong
baobaorong's picture
Hello, gsovak!

Hello, gsovak!
Do you know how many cells and how much solution they used for one sample?

Thanks!

Xin