Lipid Extraction The prediction made was that adipose tissue most likely had the highest lipid content and skeletal muscle had the lowest lipid content when comparing liver, muscle, and adipose tissue. The percent lipid was found to be 30% in liver tissue, 70% in muscle tissue, and 90% in adipose tissue in the samples collected.
Protein Extraction and Quantification The absorbance vs concentration curve prepared from the Bradford Protein Assay standards was found to have a linear regression of y = 2.2184x + 0.0626 as shown in Figure 1. The absorbances of the liver, muscle, and adipose tissue were used to determine the protein concentrations of the samples. The percent protein was found to be 15.64% in liver tissue, 4.13% in muscle …show more content…
The plot of relative mobility versus the log of molecular mass of the proteins had a linear regression of y = -1.925 + 2.25 and is shown in Figure 5. This was then used to determine the molecular mass of alkaline phosphatase from the observed relative mobility. The molecular mass was found to be 77.264 kDa.
FIGURE 5. Relative Mobility vs log of Molecular Mass. This plot shows the relative mobility of each protein on the ladder versus the log of molecular weight had a linear regression of y = -1.925x + 2.25.
DISCUSSION
When comparing the lipid content in liver, skeletal muscle, and adipose tissue, the prediction made was that adipose tissue most likely had the highest lipid content and skeletal muscle had the lowest lipid content when comparing liver, muscle, and adipose tissue. The results showed that adipose tissue had the highest lipid content while liver tissue had the lowest lipid content. The prediction made of high lipid content in the adipose tissue was supported by the results. The prediction made was that skeletal muscle would have the lowest lipid content, however the results suggest that liver tissue has the lower lipid content. Adipose tissue is a major sit of storage for lipids in the form of triglycerides. The liver oxidizes lipids for energy, so lipid content is low. High lipid content in skeletal muscle …show more content…
Cells also consist of small molecules and water, which makes up about 65% by mass of an average human cell (4).
There were more bands present for each tissue type on the SDS-PAGE gel than the Simple-PAGE gel. This is due to the fact that SDS denatures proteins, allowing them to be unfolded and individual polypeptide chains. The Simple-PAGE gel does not denature the proteins, and thus proteins are run in a folded and potentially active shape. An example of this is that alkaline phosphatase activity was detected in the Sigma Red stained Simple-PAGE gel but nor the SDS-PAGE gel. Thus AP had the proper folding to be active in the Simple-PAGE gel.
Based on the relative mobility of alkaline phosphatase, the molar mass of AP was found to be 77.264 kDa. Alkaline phosphatase (AP) has a molecular mass of 89 kDa (2). The observed value is lower than the actual value. This may be due to an error while measuring the relative mobility of AP on the gel. However, the observed and actual values are similar showing that the method of protein detection by comparing bands to a protein ladder is effective for