Two additional samples were made; one was blank and the other was for the specific homogenate sample. Then 3 micrometers of Bradford assay reagent, which indicates the amount of protein present by color, was added to all samples. The spectrophotometer was zeroed at 595 NM. A standard curve was made with the different absorbency and concentrations. After the linear equation was formed, the unknown sample concentration was determined using the standard curve equation. A Gel Electrophoresis was used to perform a qualitative analysis.
The use of 5 micrometers of the homogenate was heated to 80 degrees Celsius. Then the homogenate was transferred to a 2-micrometer-protein gel sample buffer. Samples loaded on to the gel was run at 100 v and stained with comatose blue; observations were made next lab. (Gladdening 2014) The amount of glucose and glycerol were determined from the homogenate sample as well. Samples A(+) and B(-) were used to determine glucose. Sample A had OMG/ml maliciousness’s in . MM citrate buffer, pH 5. 0 and was incubated at 370 Celsius for two hours to allow for the enzyme to digest the glycogen.
This concentration was multiplied by two to account for the dilution. The liver had the most glycogen concentration (2. 499 MGM/ml) compared to the heart(0. 684 MGM/ml) and kidney(0. 904 MGM/ml)(Figure 4) 011176000 Figure 4. Concentration of glycogen is shown in the heart, kidney, and liver homogenates of boss Taurus. Errors bars were created from 5 readings for heart, 6 readings for liver, and 7 readings for kidney. Free glucose were more abundant in the liver with a concentration of 1. 451 MGM/ml compared to the heart (. 3129 g/ml) and kidney (. Magma/ml).
Free glucose concentrations were determined by using the non-enzyme treated sample. Using the standard curve equation, concentration was found and multiplied by 2 to account for dilution. Figure 5 Free glucose concentrations of heart, kidney, and liver homogenates of boss Taurus. Errors bars were created from 5 readings for heart, 6 readings for liver, and 7 readings for kidney. Figure 4 and 5 both indicated a similar pattern. The liver for glycogen had a concentration of 2. 499 MGM/ml and a free glucose concentration of 1. 451 MGM/ml. When compared to the heart and liver, the heart