Application

This kit can be used for detection of Cholinesterase (CHE) activity in animal tissue, serum (plasma), whole blood and cultured cells, and cell culture supernatant. 

Detection significance

CHE is mainly produced in liver cells and used to estimate the reserve capacity of liver and     prognosis of liver diseases. The CHE activity usually reduced during liver diseases, and the degree of reduction was related to the severity of the disease. CHE activity decreased obviously during compensated cirrhosis, which was especially significant in coma period, while there was no change of CHE activity in obstructive jaundice. CHE activity decreased obviously during Amiba's liver abscess and can return to normal in a short time after treatment, therefore, it has certain help for the research of Amiba's disease. In some extra hepatic diseases, such as systemic malnutrition, severe anemia, cancer, advanced schistosomiasis, sepsis, severe tuberculosis (especially renal tuberculosis or serous tuberculosis), CHE activity may decrease moderately or severely, and the increasing of this enzyme can be observed in nephritic syndrome, hyperthyroidism, diabetes, high blood pressure, high blood fat.

Detection principle

CHE in serum and tissues can hydrolyze acetylcholine into bilineurine and acetic acid. The remained undecomposed acetylcholine will react with hydroxylamine and form acetohydroxamic acid, which can then react with Fe ion in acid solution and form brown color compound. The activity of CHE can be calculated according to the shade of color reaction.

Experimental instrument

Test tube, Low-speed centrifuge, Micropipettor, Vortex mixer, 37℃ water bath, Spectrophotometer (520 nm)

Sample pretreatment

1.       Serum (plasma):

Detect the serum sample directly.

2.       Tissue:

Weigh the tissue accurately, add 9 times of normal saline at the ratio of Weight (g): Volume (mL) = 1:9. Make the mechanical homogenization in ice water bath to prepare 10% homogenate. Centrifuge at 2500 rpm for 10 min and take the supernatant for detection. Meanwhile, determine the protein concentration of supernatant (E-BC-K168-S, E-BC-K165-S).

3.       Whole blood:

Bottom up the heparin anticoagulated whole blood to make it mix fully, then take 0.1 mL of the whole blood and add 0.4 mL of pre-cooled double distilled water. Mix fully for 1 min and stand for 15 min until the prepared hemolysis is transparent when observing under light.

Operation steps

1.       Operation table for serum (plasma) and tissue samples:

Reagent 1 (mL)

Reagent 4 (mL)

2.       Operation table for whole blood samples:

Reagent 1 (mL)

Reagent 4 (mL)

Notes

1.       This kit is for research use only.

2.       Instructions should be followed strictly, changes of operation may result in unreliable results.

3.       The validity of kit is 6 months.

4.       Do not use components from different batches of kit.