Ldh Flashcards
Major tissue source of LDH
Heart
Erythrocytes
Liver
Kidney
LDH is found in small amount in
Smooth muscle
Brain
Lungs
Elevated surem levels of LDH id observed in (?)
Myocardial infarction
Liver disease
Renal disease
Certain forms of anemia
Malignant diseases
Progressive muscle dystrophy
Lactate and pyruvate offers a number of advantages such as
Better reagent stability
Elemintaion of pre-incubation in the procedure
Rate of reaction being linear to a wide range
Less contamination of NAD with inhibiting products
Catalyzes the oxidation of lactate to pyruvate in the presence of NAD
Lactate dehydrogenase
Wavelength of LDH
340 nm
Forward reaction pH of LDH
8.3 - 8.9
Reverse reaction pH of LDH
7.1 - 7.4
Advantages of reverse reaction
Small volume
Short incubation time
Disadvantages of reverse reaction
Prone to substrate exhaustion
Loss of linearity
LDH does not act on (?) and only (?) acts as the coenzyme
D-lactate
NAD
In lactate dehydrogenase reaction, the (?) formed will proceed to the (?) pathway for generation of energy
Pyruvate
Embden-meyerhoff pathway
With the oxidation of lactate to pyruvate, pyruvate and NADH are formed
Colorimetric method based on Wacker
In the Colorimetric method based on wacker, a mixture of (1) and (2) reacts with NADH to produce (3) colored compound
1 Phenazine methosulfate
2 nitroblue tetrazolium
3 Blue
conversion of pyruvate to lactate by LDH with the presence of NADH
Wroblewski and La due
pH 7.4 - 7.8
The rate of reverse reaction in wrobleski and la due is (?) times faster than forward reaction allowing (advantages)
three
smaller volume samples and Shorter reaction time
Methods for separation and quantitation of LDH isoenzyme
1 Electrophoresis
2 Column chromatography
3 Immuno-inhibition
4 Babson’s method
5 Kaplan and Berger
6 using alpha-hydroxybutyrate as a substrate
Separates isoenzyme based on their anodal rate of migration in an electrophoretic field
Electrophoresis
(LDH Isoenzyme)
Migrates the fastest in electrophoresis
LDH1
(LDH Isoenzyme)
Migrates the lowest in electrophoresis
LDH6