Free cholesterol produces 4-cholestenone and hydrogen peroxide under the oxidation of cholesterol oxidase. In the presence of 4-aminoamylpyridine and phenol, peroxidase catalyze hydrogen peroxide to form red quinone compounds of benzoquinone imine phenizone. The color depth of the generated quinones is directly proportional to the cholesterol content.
Performance characteristics
| Synonyms |
FC |
| Sample type |
Serum, plasma, tissue |
| Sensitivity |
0.07 mmol/L |
| Detection range |
0.07-24 mmol/L |
| Detection method |
Colorimetric method |
| Assay type |
Quantitative |
| Assay time |
20 min |
| Precision |
Average inter-assay CV: 5.200%Average intra-assay CV: 1.900% |
| Other instruments required |
Micropipettor, Incubator, Vortex mixer, Centrifuge |
| Other reagents required |
Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4), Isopropanol(AR) |
| 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 (0.07-24 mmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
Human plasma
|
1
|
|
Human serum
|
1
|
|
Rat serum
|
1
|
|
Mouse serum
|
1
|
|
Rabbit serum
|
1
|
|
10% Rat kidney tissue homogenate
|
1
|
Note: The diluent is normal saline (0.85% NaCl) or PBS (0.01 M, pH 7.4).
