Enzymes assays Flashcards
Many diseases that cause tissue damage / injury result in ——release of intracellular enzymes into the —-.
inc.
plasma
What is Macro-amylase-emia?
Macromolecules of amylase (aggregates with Ig - LDH, ALP, CK) = rate of clearance reduced = Amylase Higher than normal
Which of the following enzymes is normally detected in urine, and why?
Amylase bc the other enzymes are too large to cross the glomerulus
pH range of blood
7.35-7.45
During purification process of enzymes, the assay must be __ because the ___ of most enzymes are very low.
highly sensitive
specific activities
When measuring enzyme activity, you add acidic 2,4-dinitrophenylhydrazine, why?
(after 30min of assay) Denature enzymes = stop activity = can be measured at 505nm = detect end point
Equation to finding [ ] of test =
[ Abs(test) / Abs(Std) ] x C(std) *NOTE: equ. comes from A(test) C(test) \_\_\_\_\_ = \_\_\_\_\_ A(Std) C(Std)
define product inhibition
too much product can inhibit the enzyme bc product can bind again to active site & prevent new substrate reacting
Describe coupled enzyme assays(*cont)
if reactant or product is colourless*
State two factors that can raise plasma ALP activity and they are not diseases? Explain your answer.
- Age, sex, race/ethinicity, physiological conditions
* 3rd trimester pregnancy: 30 – 125 U/L
in protein electrophoresis what proteins are found on the gamma region (near the anode)?
- CRP
- Ig*
- Fibrinogen
in protein electrophoresis what proteins are found on the beta region?
- transferrin
- LDL
in protein electrophoresis what proteins are found on the alpha-2 region?
- Haptoglobin
- HDL
Describe the order of zones in protein electrophoresis from cathode to anode.
(neg) Albumin - alpha1 - alpha2 - beta - gamma (pos)
Enzymes can give information on
- Presence of disease
- Which organ(s) is diseased
- Extent / severity of the disease – more cell damaged means more leaked enzymes … severe disease
- Time course of the disease
