1. For tissue, cells and cell culture supernatant sample:
1) Enzymatic reaction (Reagent 1 application solution are pre-warmed in advance at 37℃)
| Non-enzyme tube | Enzyme tube |
1 mmol/L GSH (mL) | 0.2 | 0.2 |
Sample (mL) |
| 0.2 |
Pre-heat the tubes at 37℃ water bath for 5 min. Preheat Reagent 1 application solution at 37℃ for 5 min at the same time. | ||
Reagent 1 application solution (mL) | 0.1 | 0.1 |
React at 37℃ water bath for 5 min accurately. | ||
Reagent 2 (mL) | 2 | 2 |
Sample (mL) | 0.2 |
|
Mix fully and centrifuge at 3100 g for 10 min, then take 1 mL of supernatant for chromogenic reaction.
2) Chromogenic reaction
| Blank tube | Standard tube | Non-enzyme tube | Enzyme tube |
GSH standard application solution (mL) | 1 |
|
|
|
20 μmol/L GSH standard solution (mL) |
| 1 |
|
|
Supernatant (mL) |
|
| 1 | 1 |
Reagent 3 application solution (mL) | 1 | 1 | 1 | 1 |
Reagent 4 (mL) | 0.25 | 0.25 | 0.25 | 0.25 |
Reagent 5 application solution (mL) | 0.05 | 0.05 | 0.05 | 0.05 |
Mix fully and stand for 15 min at room temperature. Set to zero with double-distilled water and measure the OD values of each tube at 412 nm with 1 cm optical path quartz cuvette.
2. For serum/plasma sample:
1) Enzymatic reaction (Reagent 1 application solution are pre-warmed in advance at 37℃)
| Non-enzyme tube | Enzyme tube |
1mmol/L GSH (mL) | 0.2 | 0.2 |
Sample (mL) |
| 0.1 |
Pre-heat the tubes at 37℃ water bath for 5 min. Preheat Reagent 1 application solution at 37℃ for 5 min at the same time. | ||
Reagent 1 application solution (mL) | 0.1 | 0.1 |
React at 37℃ water bath for 5 min accurately. | ||
Reagent 2 (mL) | 2 | 2 |
Sample (mL) | 0.1 |
|
Mix fully and centrifuge at 3100 g for 10 min, then take 1 mL of supernatant for chromogenic reaction.
2) Chromogenic reaction
| Blank tube | Standard tube | Non-enzyme tube | Enzyme tube |
GSH standard application solution (mL) | 1 |
|
|
|
20 μmol/L GSH standard solution (mL) |
| 1 |
|
|
Supernatant (mL) |
|
| 1 | 1 |
Reagent 3 application solution (mL) | 1 | 1 | 1 | 1 |
Reagent 4 (mL) | 0.25 | 0.25 | 0.25 | 0.25 |
Reagent 5 application solution (mL) | 0.05 | 0.05 | 0.05 | 0.05 |
Mix fully and stand for 15 min at room temperature. Set to zero with double-distilled water and measure the OD values of each tube at 412 nm with 1 cm optical path quartz cuvette.
| Detection range | 12.65-387 U | Average inter-assay CV | 9.300000000000001% |
| Sensitivity | 12.65 U | Average intra-assay CV | 4.9% |
| Average recovery rate | 105% |
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