Total Cholesterol And Cholesteryl Ester Fluorometric Assay Kit

Cat.No.:E-BC-F032
Detection instrument: Fluorescence microplate reader (Ex/Em=535 nm/590 nm)

Detection method: Fluorescence method

Detection principle

Total Cholesterol (TC) includes free cholesterol (FC) and cholesteryl esters (CE). Cholesterol ester can be hydrolyzed by cholesterol esterase to produce cholesterol and free fatty acid. Cholesterol is oxidized by cholesterol oxidase to produce △4-cholestenone and hydrogen peroxide. In the presence of the enzyme and probe, hydrogen peroxide can be catalyzed to produce the fluorescence substrate. The fluorescence intensity at the excitation wavelength of 535 nm and emission wavelength of 590 nm is proportional to the cholesterol concentration.

Performance characteristics

Sample type	Serum, plasma, animal tissue, cells
Sensitivity	0.12 μmol/L
Detection range	0.12-30 μmol/L
Detection method	Fluorescence method
Assay type	Quantitative
Assay time	20 min
Precision	Average inter-assay CV: 7.300%Average intra-assay CV: 1.700%
Other instruments required	Micropipettor, Vortex mixer, Centrifuge
Storage	-20℃
Valid period	12 months

Both avasimibe treatment and SOAT1 knockout can reduce the TC level in GC cells. (P<0.001)

T Zhu et al investigate the effects of SOAT1 and Avasimibe in gastric cancer metastasis. Total cholesterol and cholesteryl ester (TC) levels in GC cell was determined using TC fluorometric assay kit (E-BC-F032).

Dilution of sample

It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (0.12-30 μmol/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type	Dilution factor
Human serum	100-300
Rat serum	100-300
Mouse plasma	100-300
Rabbit serum	100-300
10% Rat liver tissue homogenate	50-150
10% Mouse kidney tissue homogenate	50-200
10% Rat brain tissue homogenate	200-400
10% Rat spleen tissue homogenate	50-200
Jukat cells	20-50