ALP Flashcards
What is the known function of ALP?
Lipid transfer in the intestine and bone formation
This hydrolyzes naturally occurring substrate.
ALP
What is the optimum pH of ALP?
10.3
What is the systematic name of ALP?
E.C. 3.1.3.1
What are the major isoenzymes of ALP?
- Placental Isoenzyme
- Intestinal Isoenzyme
- Liver Isoenzyme
- Bone Isoenzyme
What method is used for ALP measurement?
Bowers and McComb
This method uses p-NPP substrate. Thus, chelators falsely lower activity. First, enzyme activity increases slightly on storage and is relatively stable at 4’C up to a week.
Bowers and McComb
In storage, at what rate does enzyme activity increase per day?
2%
Under Bowers and McComb, enzyme activity will remain stable at what temperature?
4’C up to a week.
If the sample is frozen. What should we do?
- Thaw
- Store the sample at RT for 18 - 24hrs before testing.
If the sample is frozen. What should we do?
- Thaw
- Store the sample at RT for 18 - 24hrs before testing.
ALP reaction only takes place in the presence of cofactors (disulfide ions). What are these?
- Cobalt
- Magnesium
- Manganese
- Zinc
This cofactor serves as a constituent metal ions and must combine with ALP reaction to take place because without this factor ALP reaction would be incomplete.
Zinc
What are the Inhibitors of ALP?
- Phosphates
- Borate
- Oxalate
- Cyanide ions
If the anticoagulant contains these inhibitors, it should never be used in measuring ALP. What are these?
Borate and Oxalate
What isoenzymes do this inhibit?
A. Phenylalanine
B. Levamisole
A. Intestinal and Placental
B. Liver and Bone
This separation of enzymes is differentiated based on heat stability.
Heat fractionation.
In heat fractionation, at what degrees is the enzyme exposed to?
56’C for 10 minutes
What isoenzyme is separated in Heat Fractionation?
All except Intestinal Isoenzyme
In electrophoresis, it is exposed to what enzyme?
Neuraminidase
In electrophoresis, what are we going to do with the neuraminidase?
For 15 minutes, pretreat the neuraminidase + serum @ 37’C
What are the substrates used in Electrophoresis?
- Agarose gel
- Polyacrylamide gel
- Cellulose acetate
This improves separation of bone and liver form. Differences in their CHO side chains.
Use of enzymes or lectins
What overlaps with bone isoenzyme?
Placental Isoenzyme
In chronological order, what ALP isoenzyme is the fastest?
Liver > Bone > Placental > Intestinal Isoenzyme
What are the methods for the separation of Isoenzymes?
- Inhibition
- Heat fractionation
- Electrophoresis
- Use of enzymes or lectins
ALP elevation happens if there is?
- Obstructive hepatic disorders
- Paget’s disease
- B1x
- DM, renal failure and cirrhosis
- ALP isoenzymes in cancer patients
In extensive obstruction, there will be ________increase in the ULN of ALP.
3 FOLDS
This refers to the bone ALP isoform in the serum of dialysis patients.
B1x
What ALP isoenzyme will increase in DM, renal failure and Cirrhosis?
Increased Intestinal ALP
Physiological elevation are seen during?
- Pregnancy
- Infant and Children
Physiological elevation are seen during?
- Pregnancy
- Infant and Children
In infants and Children, how many times is the elevation?
10x
What ALP isoenzyme is present in Infants and Children?
Liver and Bone isoenzymes
This isoenzyme is present to patients with a particular type of lung CA.
Regan Isoenzyme
This isoenzyme is present to patients that have adenocarcinoma
Nagao Isoenzyme
This is present to patients that have hepatoma.
Kasahara isoenzyme
