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Glycolate Oxidase Activity Assay Kit– MSE Supplies LLC

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Glycolate Oxidase Activity Assay Kit

SKU: E-BC-K692-S-100

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Glycolate Oxidase Activity Assay Kit

SKU # E-BC-K692-S
Detection Instrument Spectrophotometer (324 nm)
Detection method Colorimetric method


Product Details

Properties

Sample type Plant tissue
Sensitivity 0.3 U/mL
Detection range 0.3-350 U/mL
Detection Method Colorimetric method
Assay type Enzyme Activity
Assay time 30 min
Precision Average inter-assay CV: 8.200% | Average intra-assay CV: 1.100%
Other instruments required Test tube, Micropipettor, Vortex mixer, Optical path cuvette (1 mL of volume, 1 cm optical diameter)
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 (0.3-350 U/mL).

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

Sample type Dilution factor
10% Oxalis corniculata tissue homogenate 1
10% Ginkgo biloba tissue homogenate 1
10% Cactus tissue homogenate 1
10% Bamboo leaf tissue homogenate 1
10% Osmanthus fragrans leaf tissue homogenate 4-8
10% Camphor trees leaf tissue homogenate 3-5

 

Note: The diluent is reagent 1.

 

Detection Principle

Glycolate oxidase catalyzes sodium glycolate substrate to form glyoxylic acid, which reacts with phenylhydrazine hydrochloride to form phenylhydrazone glyoxalate. The substance has an absorption peak at 324 nm, and its OD value is proportional to the concentration of phenylhydrazone glyoxalate in a certain range, and the amount of phenylhydrazone generated reflects the activity of glycolate oxidase.

 

Kit Components & Storage

Item Component Size 1
(50 assays)
Size 2
(100 assays)
Storage
Reagent 1 Extraction Solution 60 mL × 1 vial 60 mL × 2 vials 2-8℃, 12 months
Reagent 2 Buffer Solution 45 mL × 1 vial 45 mL × 2 vial 2-8℃, 12 months
shading light
Reagent 3 Chromogenic Agent Powder × 1 vial Powder × 1 vial 2-8℃, 12 months, shading light
Reagent 4 Substrate 6 mL × 1 vial 12 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 cactus tissue 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/mL) 5.80 85.50 179.00
%CV 1.3 1.2 0.8


Inter-assay Precision

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

Parameters Sample 1 Sample 2 Sample 3
Mean (U/mL) 5.80 85.50 179.00
%CV 8.0 8.1 8.5

 

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%.

 

Sensitivity

The analytical sensitivity of the assay is 0.3 U/mL. 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.