Arylesterase Assay Kit

FILES


- Package Insert Of Arylesterase Assay Kit.pdf

- Related_Articles.pdf

 

Paraoxonase-1 (PON1) is an high density
lipoprotein (HDL)-associated enzyme with
antioxidant and antiatherogenic functions,
protecting lipoproteins against oxidative
modification. It also catalyzes the hydrolysis of
organophosphates such as paraoxon and
aromatic carboxylic acid esters of fatty acids. It
has been shown that serum paraoxonase
activity decrease in diabetes mellitus, coronary
artery disease, hypercholesterolaemia, iron
deficiency anemia, hepatitis, cirrhosis, prostate
cancer, tuberculosis and inflammations.


Principle of Assay
PON1, present in the sample, hydrolyses
phenyl acetate to its products which are phenol
and acetic acid. The produced phenol is
colorimetrically measured via oxidative
coupling with 4-aminoantipyrine and potassium
ferricyanide. Nonenzymatic hydrolysis of
phenyl acetate was subtracted from the total
rate of hydrolysis. The molar absorptivity of
colored complex is 4000 M-1 cm-1 and one
unit of arylesterase activity is equal to 1 mmol
of phenyl acetate hydrolyzed per liter per
minute at 37oC.


Storage Conditions
This kit should be stored at 4oC.


Samples
Blood serum, heparinized plasma, semen
plasma, cell lysates and tissue homogenates
can be used as sample.

Components
All reagents are ready to use.

  • Diluent Solution 50 ml
  • Reagent 1
  • Deionized Water (Any deionized water can be used)
  • Reagent 2 10 ml
  • Reagent 3 15 ml

Procedure
This assay requires predilution of sample, use
the diluent which is inside of the box for this
procedure.
Dilution ratio 1/100. (Sample/ Diluent)
Diluted Sample 3 ul
Reagent 1 260 ul
Reagent 2 10 ul
Reagent 3 80 ul
Primary Wavelength: 548 nm. , Secondary
Wavelength 700 nm.
Method : End Point - Bichromatic


Measuring Points: First absorbance must be taken after mixing sample and R1. Last
absorbance must be taken after adding and
mixing R2 and R3 when the curve reaches a
plateu. (Approximately 3-4 minutes after mixing)
Calibration Type : Linear
Factor : 1316


Manual measurement
In manual working, the volumes of the sample
and the reagents are increased at same ratio
according to the above values.


Interference and stability
Calcium chelators such as EDTA and citrate
inhibited arylesterase activity. Heparin,
hemolysis and bilirubin did not interfere the the
assay. No significant difference was observed
between fresh and non fresh serum
arylesterase activities.


 
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