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Glutamate Synthase (GOGAT) Activity Assay Kit– MSE Supplies LLC

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Glutamate Synthase (GOGAT) Activity Assay Kit

SKU: E-BC-K659-M-96

  • $ 32395
  • Save $ 5700



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..