Glutamate Synthase (GOGAT) Activity Assay Kit

Glutamate Synthase (GOGAT) Activity Assay Kit

SKU #	E-BC-K659-M
Detection Instrument	Microplate reader (330-350 nm, optimum wavelength: 340 nm)
Detection method	Colorimetric method

Product Details

Properties

Synonyms	GOGAT
Sample type	Serum, plant and animal tissue
Sensitivity	8.84 U/L
Detection range	8.84–321.05 U/L
Detection Method	Colorimetric method
Assay type	Enzyme Activity
Assay time	15 min
Precision	Average inter-assay CV: 5.800% | Average intra-assay CV: 2.500%
Other instruments required	Centrifuge
Storage	-20℃
Valid period	12 months

Dilution of Sample

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

Sample type	Dilution factor
10% Rat kidney tissue homogenate	2-4
10% Rat liver tissue homogenate	2-4
10% Rat heart tissue homogenate	2-4
10% Mouse liver tissue homogenate	2-4
Bovine serum	1
10% pleurotus cornucopiae tissue homogenate	1
10% beech mushroom tissue homogenate	1

 

Note: The diluent is extraction solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.

 

Detection Principle

GOGAT catalysis the reaction that transfer the amino from glutamine to a-KG to form two molecules of glutamic acid using NADH as the electron donor. The decreasing rate of NADH that can be measured at 340 nm can reflect the activity of GOGAT.

 

Kit Components & Storage

Item	Component	Size 1(48 T)	Size 2(96 T)	Storage
Reagent 1	Extraction Solution	50 mL × 1 vial	50 mL × 2 vials	-20℃, 12 months
Reagent 2	Buffer Solution	16 mL × 1 vial	26 mL × 1 vial	-20℃, 12 months
Reagent 3	Substrate A	Powder × 2 vials	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 4	Substrate B	Powder × 2 vials	Powder × 2 vials	-20℃, 12 months, shading light
Reagent 5	Chromogenic Agent	Powder × 2 vials	Powder × 2 vials	-20℃, 12 months, shading light
	UV 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 (U/L)	13.50	84.50	167.00
%CV	2.9	2.4	2.2

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 (U/L)	13.50	84.50	167.00
%CV	5.7	5.4	6.3

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 104%.

	Sample 1	Sample 2	Sample 3
Expected Conc. (U/L)	56.7	148	252
Observed Conc. (U/L)	56.1	155.4	272.2
Recovery rate (%)	99	105	108

 

Sensitivity

The analytical sensitivity of the assay is 8.84 U/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..