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Total Amino Acids (T-AA) Colorimetric Assay Kit– MSE Supplies LLC

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Total Amino Acids (T-AA) Colorimetric Assay Kit

SKU: E-BC-K055-M-96

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Total Amino Acids (T-AA) Colorimetric Assay Kit

SKU # E-BC-K055-M
Detection Instrument Microplate reader (640-660 nm, optimum wavelength: 650 nm)
Detection Method Colorimetric method


Product Details

Properties

Synonyms T-AA
Sample Type Serum, plasma, urine, tissue
Sensitivity 3.03 mmol/L
Detection Range 3.64-100 mmol/L
Detection Method Colorimetric method
Assay type Quantitative
Assay time 40 min
Precision Average inter-assay CV: 6.500% | Average intra-assay CV: 4%
Other instruments required Test tube, Micropipettor, Vortex mixer, Centrifuge
Other reagents required Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4)
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 (3.64-100 mmol/L).

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

Sample type Dilution factor
Human serum 1
Human urine 1
Rat plasma 1
Porcine serum 1
10% Rat heart tissue homogenate 1
10% Rat liver tissue homogenate 1
10% Mouse liver homogenate 1
10% Epipremnum aureum leaf tissue homogenate 1

 

Note: The diluent is reagent 5.

 

Detection Principle

Copper ions can complex with various amino acids to produce blue-green complex compound, and the depth of color is proportional to the content of total amino acids at a specific wavelength. T-AA content can be calculated with the absorbance at 650 nm.

 

Kit Components & Storage

Item Component Size 1(48 T) Size 2(96 T) Storage
Reagent 1 Powder A Powder × 1 vial Powder × 1 vial 2-8℃, 12 months
Reagent 2 Acid Reagent 0.4 mL × 1 vial 0.8 mL × 1 vial 2-8℃, 12 months
Reagent 3 Powder B Powder × 1 vial Powder × 1 vial 2-8°C, 12 months
Reagent 4 Powder C Powder × 1 vial Powder × 1 vial 2-8℃, 12 months
Reagent 5 Protein Precipitator 8 mL ×1 vial 15 mL ×1 vial 2-8℃, 12 months
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) 10.50 42.60 88.10
%CV 4.2 3.8 4.0


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 (mmol/L) 10.50 42.60 88.10
%CV 7.0 6.3 6.2


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%.


Standard 1 Standard 2 Standard 3
Expected Conc. (mmol/L) 14.5 45 73.2
Observed Conc. (mmol/L) 14.6 44.6 77.6
Recovery rate (%) 101 99 106


Sensitivity

The analytical sensitivity of the assay is 3.03 mmol/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 (μmol/L) 0 10 20 40 50 60 80 100
Average OD 0.070 0.086 0.098 0.126 0.141 0.156 0.186 0.214
Absoluted OD 0.000 0.016 0.028 0.056 0.071 0.086 0.116 0.144