Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
靶点:
Glutathione
Synoyms:
GSH;Glutathione; L-Reduced Glutathione
检测方法:
竞争抑制(定量分析)
灵敏度:
less than 0.52ug/mL
检测时间:
2 hours
特异性:
This assay has high sensitivity and excellent specificity for detection of Glutathione (GSH). No significant cross-reactivity or interference between Glutathione (GSH) and analogues was observed.
精密度:
Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Glutathione (GSH) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Glutathione (GSH) were tested on 3 different plates, 8 replicates in each plate. CV(%) = SD/meanX100; Intra-Assay: CV<10%; Inter-Assay: CV<12%.
稳定性:
The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition. To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.
检测原理:
This assay employs the competitive inhibition enzyme immunoassay technique. A monoclonal antibody specific to Glutathione (GSH) has been pre-coated onto a microplate. A competitive inhibition reaction is launched between biotin labeled Glutathione (GSH) and unlabeled Glutathione (GSH) (Standards or samples) with the pre-coated antibody specific to Glutathione (GSH). After incubation the unbound conjugate is washed off. Next, avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. The amount of bound HRP conjugate is reverse proportional to the concentration of Glutathione (GSH) in the sample. After addition of the substrate solution, the intensity of color developed is reverse proportional to the concentration of Glutathione (GSH) in the sample.
实验流程:
1. Prepare all reagents, samples and standards;
2. Add 50ul standard or sample to each well.
And then add 50ul prepared Detection Reagent A immediately.
Shake and mix. Incubate 1 hour at 37℃;
3. Aspirate and wash 3 times;
4. Add 100ul prepared Detection Reagent B. Incubate 30 minutes at 37℃;
5. Aspirate and wash 5 times;
6. Add 90ul Substrate Solution. Incubate 10-20 minutes at 37℃;
7. Add 50ul Stop Solution. Read at 450 nm immediately.