An Experimental and Statistical Analysis of SDH activity as compared in Liver, Kidney, and Heart Homogenates of the Bos taurus
Methods
The three tissues being analyzed in this experiment, those of the kidney, heart and liver, were taken from the animal Bos taurus. The tissue homogenates used were made by adding 1 gram of tissue to 20 ml of sucrose phosphate buffer. The buffer was composed of 250 mM sucrose, 50 mM NaPO4, with a pH of 7.4. This mixture was homogenized with a high-speed blender for 3 pulses for a duration of 10 seconds each. The homogenates were then filtered through a cheesecloth and stored at -70° C.
The homogenate used in this experiment was liver homogenate. Five different samples and a blank…show more content… Figure 2 shows the effect of inhibitor on SDH activity, as SDH activity decreased tremendously in inhibited homogenates, as opposed to uninhibited homogenates. In a comparison of the effect of 2 mM malonate between kidney and liver, there was no significant increase in SDH activity seen (p = 0.926741). This information can be seen in the attached statistical data sheet.
Figure 2 Graph depicting dose-response to differing concentrations of malonate, an inhibitor of SDH activity. Error bars represent standard deviation as calculated from the mean. Discussion
Two hypotheses were made regarding before this experiment was carried out. The hypotheses were as follows:
Hypothesis 1: Kidney homogenates would display the highest level of SDH activity, as it has many mitochondria and is involved in many metabolic functions.
Hypothesis 2: Malonate should bind to and inhibit SDH, leading to malonate’s lowering of SDH activity. The SDH from each homogenate will have different sensitivities to malonate. Sensitivities will be based on the amount of metabolic activity. The liver, kidney, and heart homogenates of the Bos Taurus were analyzed both experimentally and statistically. Through experimental evaluations of SDH activity as well as how the enzyme is affected by inhibitors shed much light on the amount of enzyme
