- Detection method: Colorimetric method
Manual
Detection principle
Total cholesterol includes free cholesterol and cholesterol esters. 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 4-aminoamylpyridine and phenol, hydrogen peroxide catalyze peroxidase to form red quinone compounds of benzoquinone imine phenizone.The color depth of the generated quinone is directly proportional to the cholesterol content. The absorbance values of the standard tube and the sample tube are measured respectively, and the cholesterol content in the sample can be calculated.
Performance characteristics
| Synonyms |
TC |
| Sample type |
Serum,plasma,hydrothorax,animal tissue |
| Sensitivity |
0.09 mmol/L |
| Detection range |
0.09-25.85 mmol/L |
| Detection method |
Colorimetric method |
| Assay type |
Quantitative |
| Assay time |
50 min |
| Precision |
Average inter-assay CV: 2.800%Average intra-assay CV: 1.100% |
| Other instruments required |
Micropipettor, Incubator, Centrifuge |
| Other reagents required |
Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4), Isopropanol |
| Storage |
2-8℃ |
| Valid period |
12 months |
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 (0.09-25.85 mmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
Human serum
|
1
|
|
Mouse serum
|
1
|
|
Rat plasma
|
1
|
|
10% Mouse liver tissue homogenate
|
1
|
|
10% Mouse kidney tissue homogenate
|
1
|
|
10% Mouse heart tissue homogenate
|
1
|
Note: The diluent of serum (plasma) is normal saline ( 0.9% NaCl ) or PBS ( 0.01 M, pH 7.4 ); The diluent of animal tissue is anhydrous ethanol.
