Reduced Glutathione (GSH) Colorimetric Assay Kit

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Reduced Glutathione (GSH) Colorimetric Assay Kit

SKU #	E-BC-K030-M
Detection Instrument	Microplate reader(405-414 nm)
Detection Method	Colorimetric method

Product Details

Properties

Synonyms	GSH
Sample Type	Serum, plasma, cell culture supernatant, tissue, cells
Sensitivity	2 μmol/L
Detection Range	2-100 μmol/L
Detection Method	Colorimetric method
Assay type	Quantitative
Assay time	25 min
Precision	Average inter-assay CV: 3.200% \| Average intra-assay CV: 1.900%
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 the pre-experiment and the detection range (2-100 μmol/L).

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

Sample type	Dilution factor
Human serum	1
Human plasma	1
10% Mouse brain tissue homogenization	1
10% Mouse liver tissue homogenization	1
Hela cell homogenization（0.999 mgprot/mL）	1
Rat serum	1
Rat plasma	1
Mouse serum	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 GSH can react with Dinitrobenzoic acid (DNTB) to form a yellow complex which can be detected by colorimetric assay at 405 nm and calculate the reduced GSH content indirectly.

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Acid Reagent	6 mL × 1 vial	12 mL × 1 vial	2-8℃, 12 months shading light
Reagent 2	Phosphate	6 mL × 1 vial	12 mL × 1 vial	2-8℃, 12 months
Reagent 3	DTNB Solution	1.5 mL × 1 vial	1.5 mL × 2 vials	2-8℃, 12 months shading light
Reagent 4	GSH Standard	3.07 mg × 1 vial	3.07 mg × 2 vials	2-8℃, 12 months
Reagent 5	GSH Standard Stock Diluent	1.5 mL× 1 vial	1.5 mL× 2 vials	2-8℃, 12 months
	Microplate	96 wells		No requirement
	Plate Sealer	2 pieces

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 (μmol/L)	5.50	26.40	64.50
%CV	2.3	2.0	1.4

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 (μmol/L)	5.50	26.40	64.50
%CV	3.5	2.9	3.2

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 96%.

	Standard 1	Standard 2	Standard 3
Expected Conc. (μmol/L)	15	45	58
Observed Conc. (μmol/L)	14.9	42.3	55.1
Recovery rate (%)	99	94	95

Sensitivity

The analytical sensitivity of the assay is 2 μmol/L GSH 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.

Standard Curve:

As the OD value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:

Concentration (μmol/L)	0	10	20	40	50	60	80	100
Average OD	0.057	0.088	0.129	0.213	0.296	0.296	0.392	0.484
Absoluted OD	0.000	0.030	0.071	0.156	0.196	0.239	0.334	0.426