Description
Glutathione Assay Kit (BA0042) (BA0042)
The Glutathione Assay Kit (SKU: BA0042) is designed to accurately measure reduced glutathione in a broad range of biological samples. Glutathione is a tripeptide of glycine, glutamic acid and cysteine, and its reduced form is vital in maintaining haemoglobin in a reduced state and protecting cells from oxidative damage. Low levels of glutathione are found in deficiencies of key enzymes involved in glutathione metabolism, such as glucose-6-phosphate dehydrogenase, glutathione synthase and glutathione reductase. The kit uses an improved 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB) method that combines deproteination and detection into a single reagent, with DTNB reacting with reduced glutathione to form a yellow product. The optical density, measured at 412nm, is directly proportional to the glutathione concentration in the sample. The optimised formulation has a long shelf life and is completely free of interference due to sample turbidity.
| Product Name: | Glutathione Assay Kit (BA0042) |
| SKU: | BA0042 |
| Detection Method: | Colorimetric |
| Detection Range: | 0.4 - 100 µM in 96-well plate |
| Sample Type: | Whole blood, plasma, serum, urine, tissue and cell extracts |
| Species Reactivity: | All |
| Assay Time: | 25 minutes |
| Kit Size: | 250 Assays |
| Equipment Required: | Microplate reader |
| Storage: | The kit is shipped at room temperature. Store all components at 4°C. |
| Shelf Life: | 12 months after receipt |
| Shipping: | Room Temperature |
This kit measures reduced glutathione in biological samples using an improved DTNB method that combines deproteination and detection into Reagent A. DTNB reacts with reduced glutathione to form a yellow product, and the optical density at 412nm is directly proportional to the glutathione concentration.
- Sensitive and accurate: linear detection range 0.4 - 100 µM in 96-well plate
- Simple and convenient: mix the DTNB reagent with sample, remove protein precipitates for proteinaceous samples, add a second reagent and read the optical density
- Low interference: amino acids and common buffers do not interfere
- Direct assays: reduced glutathione in whole blood, plasma, serum, urine, tissue and cell extracts
- Drug discovery/pharmacology: effects of drugs on glutathione metabolism
Note: The below protocol is a sample protocol. Protocols are specific to each batch/lot. For the correct instructions please follow the protocol included in your kit.
| Step | Procedure |
| 1 | Blank and Calibrator: transfer 100 µL water and 100 µL Calibrator into wells of a clear-bottom 96-well plate, and pipette 200 µL water into the Blank and Calibrator wells. |
| 2 | Samples: dilute whole blood 20-fold with water prior to assay (n = 20). Deproteination is required for blood, serum, plasma and other proteinaceous samples. Mix 120 µL sample with 120 µL Reagent A in 1.5-mL centrifuge tubes and vortex; if turbidity occurs, pellet for 5 minutes at 14,000 rpm. |
| 3 | Transfer 200 µL sample/Reagent A mixture into wells of the 96-well plate and add 100 µL Reagent B; tap plate lightly to mix. |
| 4 | Incubate 25 minutes at room temperature and read OD at 412nm. |
Subtract blank OD (water) from the Calibrator and Sample OD values. Glutathione concentration = [(ODSAMPLE − ODBLANK) / (ODCALIBRATOR − ODBLANK)] × 100 × n (µM), where n is the dilution factor (20 for blood samples). Conversion: 1 mg/dL glutathione equals 32.5 µM, 0.001% or 10 ppm.
| Component | Quantity | Storage |
| Reagent A | 30 mL | 4°C |
| Reagent B | 30 mL | 4°C |
| Calibrator (equivalent to 100 µM glutathione) | 10 mL | 4°C |
