Experiment 10 Flashcards
Method used for Total Creatine Kinase Determination?
Kinetic Method by Oliver-Rosalki, Modified by Szasz
It plays an important role in the energy-storing mechanism of tissue by catalyzing the reverse reaction
Creatine Kinase
CK is distributed in ________
Various organ
What are the highest activities in CK (increasing order)
Brain-Heart-Skeletal Muscle
The determination of CK is an aid in diagnosing:
muscular dystrophy
other diseases of the Skeletal Muscles
Myocardial infarction
hypothyroidism
renal diseases
dysfunction
A modified method was described by?
Nielson
What are the compound does Nielson added to assure maximum CK activity?
sulfhydryl compound and AMP
What are the activity does Nielson has?
CK activity and ihibit andenylate kinase activity
Optimized conditions for measuring CK were published by _____ in _____ as well as by the ________ committee on enzyme.
Szasz, 1976, Scandinavian
pH of Forward in Total CK
6.8 pH
pH of Reverse in Total CK
9.0 pH
This is a direct or forward reaction which involves the conversion of creatine and ATP to Creatine phosphate and ADP.
Tanzer-Gilbarg Method
Most Cathodal in Electrophoresis
Mitochondrial CK
Cathodal in Electrophoresis
CK-MM
It migrates to a position midway between CK-MM & CK-MB
Macro CK
Anodal in electrophoresis
CK-BB
Degree of damage to the left ventricle : SMALL MI
<10% damage
Degree of damage to the left ventricle : MEDIUM MI
10-30% damage
Degree of damage to the left ventricle : LARGE MI
> 30% damage
Protein Mass Value : SMALL MI
10-60 ug/L
Protein Mass Value : MEDIUM MI
60-225 ug/L
Protein Mass Value : LARGE MI
> 225 ug/L
% of CK-MM
96%
CK-MM:_________ ; CK-MB:________
lipoprotein, Ig
% of CK-MB
0-6%
% CK-BB
small amount
In Column Chromatography, it exchange in resin.
diethyl amino ethyl
The creatine formed in the reverse reaction is estimated fluorometrically after addition of ____?
ninhydrin
The rate of formation and utilization of ATP in the CK assay can be followed continually by bioluminiscence reaction with _____\_____.
Luciferin,luciferase
Excesses in the reaction system that may inhibit the activity of CK.
Magnesium, Chloride, Calcium, and Sulfates
Other heavy metals that also inhibits the activity of CK
zinc,copper,manganese
Excess _____ & ______ agents
iodoacetate , sulfhydryl
What are the excesses that inhibit the activity of the enzyme?
ADP(Adenosine Di-Phosphate)
Citrate
fluoride
L-thyroxine
Uric acid
Serum stability:
4hrs-Room temp
8-12hrs-4 Degree Celcius
2-3 days- Frozen
hemolyzed specimens should not be used, what are the side reactions tha may occur?
-Adenylate kinase
-Adenosine triphosphate
-Glucose-6-phosphate dehydrogenase
Where adenylate kinase occur?
Erythrocytes
Platelets
Adenylate Kinase may also occur in what tissues?
Skeletal Muscle
Brain
Liver
Heart
Kidney
Small Intestine
Linearity of Total CK
1200IU/L
serves as the cofactor for the maximum catalytic activity of enzyme
Magnesium
inhibits adenylate kinase activity
Adenosine-5’-Monophosphate
prevents oxidation of sulfhydryl group of creatine kinase
N-acetylcysteine
Chelates calcium which is an inhibitor or the enzyme’s activity
EDTA
