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Reduced Glutathione (GSH) Colorimetric Assay Kit– MSE Supplies LLC

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Reduced Glutathione (GSH) Colorimetric Assay Kit

SKU: E-BC-K030-S-100

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Reduced Glutathione (GSH) Colorimetric Assay Kit

SKU # E-BC-K030-S
Detection Instrument Spectrophotometer(420 nm)
Detection Method Colorimetric method


Product Details

Properties

Synonyms GSH
Sample Type Serum, plasma, tissue,cells
Sensitivity 0.26 mg GSH/L
Detection Range 0.26-122.8 mgGSH/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 50 min
Precision Average inter-assay CV: 2.400% | Average intra-assay CV: 1.800%
Other instruments required Micropipettor, Vortex mixer
Other reagents required Normal saline (0.9% NaCl) or 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 pre-experiment and the detection range ( 0.26-122.8 mg GSH/L).

The recommended dilution factor for different samples is as follows (for reference only):

Sample type Dilution factor
Human serum 1
Mouse serum 1
10% Mouse brain tissue homogenization 1
10% Rat liver tissue homogenization 1
Human plasma 1
Rat plasma 1
10% Carrot tissue homogenization 1
293T supernatant 1

 

Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).

 

Detection Principle

Reduced glutathione (GSH) can react with dithionitrobenzoic acid (DTNB) to produce thio-nitrobenzoic acid and glutathione disulfide. Nitromercaptobenzoic acid is a yellow compound which has the maximum absorption peak at 420 nm. The GSH content can be calculated by measuring the OD value at 420 nm.

 

Kit Components & Storage

Item Component Size 1 (50 assays) Size 2 (100 assays) Storage
Reagent 1 Acid Reagent 45 mL × 1 vial 45 mL × 2 vials 2-8℃, 12 months shading light
Reagent 2 Phosphate Powder × 1 vial Powder × 2 vials 2-8℃, 12 months
Reagent 3 DTNB Solution 1.5 mL × 1 vial 30 mL × 1 vial 2-8℃, 12 months shading light
Reagent 4 Salt Reagent Powder × 2 vials Powder × 4 vials 2-8℃, 12 months shading light
Reagent 5  GSH Standard 3.07 mg × 1 vial 3.07 mg ×2 vials 2-8℃, 12 months
Reagent 6 GSH Standard
Stock Diluent
6 mL× 1 vial 6 mL× 1 vial 2-8℃, 12 months

 

Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.

 

Technical Data:

Parameter:

Intra-assay Precision

Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).

Parameters Sample 1 Sample 2 Sample 3
Mean (mg GSH/L) 3.60 37.50 76.20
%CV 2.3 1.9 1.2


Inter-assay Precision

Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (mg GSH/L) 3.60 37.50 76.20
%CV 2.8 2.0 2.4


Recovery

Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 102%.


Sample 1 Sample 2 Sample 3
Expected Conc. (mg GSH/L) 16 45.5 85.4
Observed Conc. (mg GSH/L) 16.5 45.5 88.0
Recovery rate (%) 103 100 103


Sensitivity

The analytical sensitivity of the assay is 0.26 mg GSH/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.