1.3.MEASUREMENT OF THE ACTIVITY OF ALKALINE PHOSPHATASE (ALP) IN THE BLOOD PLASMA

Question 1

enzymes

Answer 1

are protein molecules which have catalytic effect in certain chemical reactions. they lower the energy level.

Question 2

what is needed for the function of an enzyme

Answer 2

optimal pH, substrate molecules and optimal

temperature

Question 3

ALP is released into the blood plasma when

Answer 3

the disruption of biliary epithelial cells, osteoblasts (cells that responsible for the bone formation) or liver cells (hepatocytes), and in pregnant animals from the placenta.

Question 4

what does the ALP

Answer 4

catalyze the cleavage of phosphates from ester bonds

regulating the intracellular phosphate concentration

Question 5

at what pH the ALP works the best

Answer 5

9-10

Question 6

ALP binds to

Answer 6

cell membranes (plasmamembrane, endoplasmic reticulum)

Question 7

what happens when the ALP released into the intestines?

Answer 7

it can be activated and deactivate the bacterial endotoxin

Question 8

how does ALP activate/ deactivate endotoxin?

Answer 8

by dephosphorylation

Question 9

the physiological values of ALP in horse

Answer 9

80-120 IU/l

Question 10

the physiological values of ALP in cattle

Answer 10

15-50 IU/l

Question 11

the physiological values of ALP in pig

Answer 11

20-40 IU/l

Question 12

the physiological values of ALP in sheep

Answer 12

40-50 IU/l

Question 13

the physiological values of ALP in dog

Answer 13

10-25 IU/l

Question 14

the physiological values of ALP in cat

Answer 14

10-20 IU/l

Question 15

which substrate do we add to ALP to determine its activity?

Answer 15

4-nitrophenyl-phosphate dissolved in glycine buffer pH 10.5

Question 16

which molecule will be produced after the cleavage of the ester bonds

Answer 16

4-nitrophenol

Question 17

what will be the colour of the 4-nitrophenol and how can we measure its intensity?

Answer 17

yellow. can be measured by photometry at 405 nm wavelength

Question 18

what are the materials needed for the measurement of ALP

Answer 18

 Blood plasma (3x diluted)
 Reagent (4-nitrophenyl-phosphate)
 Water bath at 37 °C
 Spectrophotometer
 Micropipette

Question 19

ALP released into the blood when

Answer 19

 obstruction of the biliary duct (e.g bile stones, parasite infection)
 damaged hepatocytes (hepatic diseases)
 pregnant animals


Question 20

how can you deactivate ALP

Answer 20

incubating the enzyme at 65 degrees for 2 minutes