An Experimental and Statistical Analysis of Sdh Activity as Compared in Liver, Kidney, and Heart Homogenates of the Bos Taurus

In the experiment we used Turnip, Hydrogen Peroxide, Distilled Water, and Guaiacol as my substances. On the first activity, Effect of Enzyme concentration of Reaction Rate for low enzyme concentration, we tested three concentrations of the turnip extract, and hydrogen peroxide. For the Turnip Extract I used 0.5 ml, 1.0 ml, and 2.0 ml. For hydrogen peroxide we used 0.1 ml, 0.2 ml, and 0.4 ml. We used a control to see the standard, and used a control for each enzyme concentration used. The control contains turnip extract and the color reagent, Guaiacol. We prepared my substrate tubes separately from the enzyme tubes. My substrate tube

After complete, continue using HCl and NaOH, but now testing potato cell, liver cell, and the buffer (one at a time)

Understanding the activity of enzymes in different muscle types can aid greatly in obtaining more information about other processes such as metabolism of the tissues (Anderson et al., 2012). There are many different methods in order to achieve this information based in two different major categories but the most convenient method is one called continuous assay. This process includes the use of a spectrophotometer to continuously monitor the assay. This method allows for an easy way to calculate the initial rate of reaction as one can establish time points easily. In the lab performed the continuous method was used in order to determine the measurement of the activity enzyme succinate dehydrogenase (SDH). SDH is most commonly found in mitochondria. It is present in many different muscle tissues including the heart, red, and white tissue. In the following lab the enzyme was tested and measured in these three muscle types in the Oncorhynchus mykiss in order to determine which type contained the highest and the lowest activity. This enzyme is involved in multiple processes such as involved in both the Krebs cycle and the electron transport chain (ETC) (Smith, 2014). Its role in

5.  What reaction would you expect when performing a negative control in the catalase assay?

2. Four unknown samples were included in the lab kit: flax seed meal, potato starch, egg whites, and dried milk. Using the results of the biochemical testing, determine which number corresponds to the correct unknown. (8 points)

The homogenates provided were made by homogenizing tissues in a sucrose phosphate buffer in a 1:20 ratio. The protein concentration in bovine cells was measured by diluting the homogenate with a 1:5 ratio; 50 microliters of homogenate and 200 microliters of water. Then 5 known protein concentration samples which were 0.4, 0.8, 1.2, 1.6, 2.0 mg/ml of bovine serum were used to determine absorbance with a spectrophotometer. Two additional samples were made; one was blank and the other was for the specific homogenate sample. Then 3 microliters of bradford assay reagent, which indicates the amount of protein present

J. Moldovan & B. Nilson, (2010), Lab 4 – Enzyme Kinetics, UBCO BIOL/BIOC 393, UBC Vista accessed Monday, November 8th, 2010.

Homogenates were provided made from liver, kidney, or heart in a 1:20 ratio with sucrose-phosphate buffer that was stored at -70° C. The tissue I tested was the liver homogenate. To qualitatively analyze proteins of the homogenate, 5µl of original liver homogenate was combined with 2 µl of protein gel sample buffer and heated in a water bath at 80° C for 10 minutes. After centrifuging the sample for 5 seconds, the contents were loaded onto a polyacrylamide protein gel set at 100V for Electrophoresis and stained Coomassie blue for an hour and results were observed the next class period a week later (Clendening).

The intention of the experiment was to measure how different pH levels affect the heart rate of daphnia. The objective of this experiment was to measure the heart rate of

There are four main factors that can impact the activity of an enzyme; Temperature, pH level, the enzymes concentration and the substrate concentration, as well as other more minor factors like the presence of any inhibitors or activators. For this experiment we focus on the temperature and pH of the enzyme.

Malonate is an inhibitor of SDH, meaning that when placed in a solution that has SDH present it will bind to the active sites of enzymes making them unable to react, ultimately slowing down the reaction. For this lab it is expected that all three homogenates (liver, kidney and heart) will all have different sensitivities to malonate due to their different functions. The heart should react the most as it needs the most energy to function followed by the liver as it needs the second most energy and then the kidney. The more energy an organ needs the more enzymes it has to produce molecules like ATP. In this experiment the three homogenates were tested in different amounts of malonate and SDH.

Acids and bases were added to the solution in order to achieve the desired pH. While there were complications leading to a shortage of data, the results pointed to a peak of enzymatic activity between 6 and 8 pH. This suggested that the hypothesis

Abstract What results would acquire when given certain changes to temperature; substrate concentration, enzyme concentration, and inhibitor existence are made considering the kinetics of a specific enzyme? While examining the results these variables significant to the enzyme kinetics will provide a understanding of the general enzyme activity. Observing the end products of each chemical reaction that included an enzyme produced a velocity of that certain enzyme involved, which is measured. These results in this experiment will show temperature change, substrate concentration change, enzyme concentration change, and existence of an inhibitor produce a positive or negative effect on the enzymes activity. Abbreviations [S]: Substrate concentration

In this experiment, NaOH was the inhibitor used to stop the enzymatic reactions. NaOH is very basic and when added to a solution, will cause a drastic increase in pH, causing denaturation of the enzyme. The amount of product formed could be calculated by placing the test tube in a spectrometer after the addition on NaOH. A spectrometer measures the absorbance of a solution, which helps compare how much of a substance is in a solution.

Six plastic cups were obtained and labeled 0.2 M, 0.4 M, 0.6 M, 0.8 M, and 1.0 M. Next, each of the six dialysis tubes were knotted on one end and filled with the sucrose samples, and then tied off. These samples were then dried by patting with a paper towel, weighed and placed into their corresponding cups. The mass of each sample was recorded in table 2 on the data sheet. Each cup was then