*(GSH)檢測試劑盒
適用生物 General，通用
*(GSH)檢測試劑盒檢測范圍 1.23-100ug/mL 靈敏度 0.46ug/mL
樣本類型 Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
實驗時長 2.5h 實驗方法 競爭抑制法 *(GSH)檢測試劑盒規格 96T
ELISA Kit for Glutathione (GSH)
FOR IN VITRO AND RESEARCH USE ONLY, NOT FOR USE IN CLINICAL DIAGNOSTIC PROCEDURES!
Organism species General
*(GSH)檢測試劑盒Sample type Serum, plasma, tissue homogenates, cell lysates, cell culture supernates and other biological fluids
Format 96-well strip plate
Assay length 2.5 hours
Detection range 1.23-100ug/mL The standard curve concentrations used for the ELISA’s were 100ug/mL, 33.33ug/mL, 11.11ug/mL, 3.7ug/mL, 1.23ug/mL
Sensitivity The minimum detectable dose of this kit is typically less than 0.46ug/mL.
Specificity
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.
Recovery
Matrices listed below were spiked with certain level of recombinant Glutathione (GSH) and the recovery rates were calculated by comparing the measured value to the expected amount of Glutathione (GSH) in samples.
Matrix Recovery range (%) Average(%)
serum(n=5) 83-105 87
EDTA plasma(n=5) 86-93 89
heparin plasma(n=5) 88-104 97
*(GSH)檢測試劑盒Precision
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%
Linearity
The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Glutathione (GSH) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.
Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-103% 90-102% 79-104% 97-105%
EDTA plasma(n=5) 94-103% 94-102% 97-104% 82-92%
heparin plasma(n=5) 94-101% 78-101% 90-97% 81-96%
Stability
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.
Reagents and materials provided
Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B 1×120μL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1
*(GSH)檢測試劑盒Assay procedure summary
1. Prepare all reagents, samples and standards;
2. Add 50μL standard or sample to each well.
And then add 50μL prepared Detection Reagent A immediay.
Shake and mix. Incubate 1 hour at 37oC;
3. Aspirate and wash 3 times;
4. Add 100μL prepared Detection Reagent B. Incubate 30 minutes at 37oC;
5. Aspirate and wash 5 times;
6. Add 90μL Substrate Solution. Incubate 15-25 minutes at 37oC;
7. Add 50μL Stop Solution. Read at 450 nm immediay.
*(GSH)檢測試劑盒Test principle
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.
