- Detection method: Colorimetric method
Manual
Detection principle
Using NAD+ as hydrogen acceptor, LDH catalyzes the conversion of both lactate and NAD+ into pyruvic acid and NADH respectively. 1-Methoxy-5-methyl phenazine methyl sulfate (PMS) transfers hydrogen from NADH to NBT which deoxidize into purple chromogenic substrate. Lactic acid content can be calculated by measuring the OD value at 530 nm.
Performance characteristics
| Sample type |
Whole blood |
| Sensitivity |
0.14 mmol/L |
| Detection range |
0.14-7.0 mmol/L |
| Detection method |
Colorimetric method |
| Assay type |
Quantitative |
| Assay time |
60 min |
| Precision |
Average inter-assay CV: 2%Average intra-assay CV: 1.600% |
| Other instruments required |
Micropipettor, Vortex mixer, Incubator, Centrifuge |
| Other reagents required |
Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| 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 (0.14-7.0 mmol/L).
The recommended dilution factor for different samples is as follows (for reference only):
|
Sample type
|
Dilution factor
|
|
Rabbit whole blood
|
1-2
|
|
Mouse whole blood
|
1-2
|
|
Rat whole blood
|
1-2
|
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
