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Proline (Pro) Colorimetric Assay Kit– MSE Supplies LLC

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Proline (Pro) Colorimetric Assay Kit

SKU: E-BC-K177-M-96

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Proline (Pro) Colorimetric Assay Kit

SKU # E-BC-K177-M
Detection Instrument Microplate reader (520 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms Pro
Sample Type Plant tissue and honey sample
Sensitivity 0.57 μg/mL
Detection Range 0.57-35 μg/mL
Detection Method Colorimetric method
Assay type Quantitative
Precision Average inter-assay CV: 4.7-10.0% | Average intra-assay CV: 3.7-5.5%
Other instruments required Vortex mixer, Water bath, Centrifuge
Other reagents required Acetic acid
Storage 2-8℃
Valid period 12 months


Detection Principle

Proline can react with acidic-ninhydrin to form stable red compound. The maximum absorption peak of the compound is at 520 nm. And the concentration of Pro can be calculated by measuring the OD value at 520 nm

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Extracting Solution 50 mL× 3 vials 60 mL× 5 vials 2-8°C, 12 months, shading light
Reagent 2 Ninhydrin 3 g × 1 vial 6 g × 1 vial 2-8°C, 12 months, shading light
Reagent 3 Acid Reagent 50 mL× 1 vial 50 mL× 2 vials 2-8°C, 12 months
Reagent 4 100 μg/mL Proline
Standard
1 mL× 1 vial 2 mL× 1 vial 2-8°C, 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 pear 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 (μg/mL) 3.50 12.60 30.20
%CV 5.5 5.2 3.7


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 (μg/mL) 3.50 12.60 30.20
%CV 10.0 4.7 7.5

 

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 100%.

Standard 1 Standard 2 Standard 3
Expected Conc. (μg/mL) 8.5 18.5 28
Observed Conc. (μg/mL) 8.8 17.0 29.1
Recovery rate (%) 104 92 104

 

Sensitivity

The analytical sensitivity of the assay is 0.57 μg/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.

 

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 (μg/L) 0 5 10 15 20 25 30 35
Average OD 0.042 0.125 0.216 0.329 0.446 0.562 0.716 0.807
Absoluted OD 0.000 0.084 0.174 0.288 0.404 0.520 0.650 0.765