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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-S-100

  • $ 30695
  • Save $ 5400



Proline (Pro) Colorimetric Assay Kit

SKU # E-BC-K177-S
Detection Instrument Spectrophotometer (520 nm)
Detection method Colorimetric method


Product Details

Properties

Synonyms Pro
Sample Type Plant tissue, honey
Sensitivity 0.17 μg/mL
Detection Range 0.17-35 μg/mL
Detection Method Colorimetric method
Assay type Quantitative
Assay time 80 min
Precision Average inter-assay CV: 5.400% | Average intra-assay CV: 4.500%
Other instruments required Water bath, Micropipettor, Vortex mixer, Balance, Centrifuge
Other reagents required Acetic acid, Methylbenzene, Absolute ethyl alcohol
Storage 2-8℃
Valid period 12 months


Images

ZH Yue et al investigate the function of Bacillus altitudinis for improving salt tolerance in wheat. Proline (Pro) level in wheat tissue was determined using Proline colorimetric assay kit (E-BC-K177-S).

The levels of proline concentration was significantly increased with salt tolerance and reversed by WR10. (**P<0.01 vs Control, ##P<0.01 vs NaCl)

 

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 (0.17-35 μg/mL).

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

Sample type Dilution factor
Green pepper tissue homogenization 1
Carrot tissue homogenization 1
Epipremnum aureum tissue homogenization 1
Lettuce leaf tissue homogenization 1
Honey 1
Cabbages  tissue homogenization 1

 

Note: The diluent is Reagent 1.

 

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
(50 Assays)
Size 2
(100 Assays)
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

 

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 honey 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) 2.80 16.40 28.50
%CV 4.7 4.4 4.4


Inter-assay Precision

Three honey 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) 2.80 16.40 28.50
%CV 5.1 5.8 5.3

 

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 105%.

Standard 1 Standard 2 Standard 3
Expected Conc. (μg/mL) 6.5 18 27.6
Observed Conc. (μg/mL) 6.9 18.5 29.3
Recovery rate (%) 106 103 106

 

Sensitivity

The analytical sensitivity of the assay is 0.17 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 30 35
Average OD 0.003 0.152 0.330 0.487 0.675 1.008 1.203
Absoluted OD 0 0.149 0.327 0.484 0.672 1.005 1.200