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Glycerol Colorimetric Assay Kit– MSE Supplies LLC

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Glycerol Colorimetric Assay Kit

SKU: E-BC-K340-M-96

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Glycerol Colorimetric Assay Kit

SKU # E-BC-K340-M
Detection Instrument Microplate reader (500-520 nm, optimum wavelength: 510 nm)
Detection method Colorimetric method


Product Details

Properties

Sample type Serum (plasma), animal tissue, cell
Sensitivity 0.01mmol/L
Detection range 0.01-1.00 mmol/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 10 min
Precision Average inter-assay CV: 8% | Average intra-assay CV: 4%
Other instruments required Water bath
Other reagents required Double distilled water, Isopropanol (AR), PBS(0.01 M,pH 7.4)
Storage 2-8℃
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% Mouse liver tissue homogenate 2
10% Mouse kidney tissue homogenate 1
10% Mouse spleen tissue homogenate 1
Cell 1

 

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

 

Detection Principle

Glycerol is transformed by enzyme to produce hydrogen peroxide, which is catalyzed by oxidase in the presence of 4-amino-antipyrprid and phenol to produce red quinones, the color of which is proportional to the content of glycerol.


Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Working Solution 12.5 mL × 1 vial 25 mL × 1 vial 2-8℃, 12 months shading light
Reagent 2 1.0 mmol/L Standard 1 mL × 1 vial 1 mL × 2 vials 2-8℃, 12 months shading light
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 (mmol/L) 0.25 0.64 0.82
%CV 4.2 3.9 3.9


Inter-assay Precision

Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.

Parameters Sample 1 Sample 2 Sample 3
Mean (mmol/L) 0.25 0.64 0.82
%CV 8.2 7.8 8.0


Sensitivity

The analytical sensitivity of the assay is 0.01mmol/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.


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
(mmol/L)
0.00 0.10 0.20 0.40 0.50 0.60 0.80 1.00
Average OD 0.072 0.122 0.175 0.293 0.340 0.394 0.505 0.601
Absoluted OD 0.000 0.050 0.103 0.221 0.269 0.323 0.433 0.530