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Cellular signaling is a fascinating process, where protein receptors, transducers, accelerators, inhibitors, and many other substances interact to produce a cellular response. Enzymes play a critical role in cellular functioning, and a single enzyme deficiency may cause our cells to fail…
Substances levels can give a lot of information to healthcare professionals, in both clinical and research settings.
Lactate dehydrogenase (LDH) is present in many human tissues, enabling cells to undergo anaerobic glycolysis; its main cellular function, conversion of pyruvate into lactate, provides the energy production cycle with more substrates. Normal and abnormal levels are now standardized, and measuring its activity in serum/plasma will help elucidate the origins of damage or disease. There are different isoenzymes of LDH, which differ in their structure and certain other properties.
In this practical work, which is divided into 3 weeks, we will first make a comparison of the absorption spectra of NAD+ and NADH, since the assay for LDH makes use of an important difference in these spectra. In addition, the linearity of the LDH assay, with respect to the amount of enzyme, will be assessed, and the limit of linearity determined. In week two, we will assess the LDH isoenzyme profile in rat serum and selected tissues, using agarose gel electrophoresis. Finally, the total LDH activity will be determined, using its natural substrate pyruvate. LDH activity will also be measured using the substrate analogue
Nicotinamide adenine dinucleotide (NAD) is a coenzyme, a molecule which aids an enzyme in the acceleration of a chemical reaction, or catalysis. ...
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