Data Analysis Experiment - Essay Example

The Experiment of Western blotting analysis for the samples of NIH/3T3 cells, whether having been treated in PDGF or not, uses the Phospho-Akt (Ser473) for the preparation of the upper blotting and Akt 9272 for the lower blotting. It has to present the results in a tabular form, the first column being PGDF, the second being AKT, and the third column being Phospho-Akt.

This experiment was meant to investigate the sensitivity of antibodies to sense the presence of and the levels of Phospho-Akt (Ser473) and Akt1 Ser473.
Hypothesis
It worked with the hypothesis that the antibodies can identify Akt2 as well as Akt3 after they phosphorylate in various residues.

Procedure
This was a time series practical activity to stimulate the cells at various durations from 0 to 120 minutes. Its independent variable was the PGDF. There are two blots to analyze, Ser473 of Akt as the upper blot (using phospho-Akt antibody) and phosphor-Akt (using Akt antibody) as the lower blot. It also involved using gels to conduct the molecular weight marker. The top band corresponded to the measurement of 76 kDa, while the middle band corresponded to 57 kDa. The lowest band thus corresponded to 47 kDa.
Findings
This experiment requires the analysis of the localization of protein for the EGFR factor with the green label as shown below:

Green label of the EGFR Factor
The experiment also requires the red labeling of endocytosis markers, which apply fluorescence microscopy as shown below:

Red label of Endocytosis Marker
The growth factor analysis shows that proteins facilitate growth at different rates under different conditions. The most relevant protein analysis procedure in this kind of experiment is fluorescent microscopy.
Hypothesis
This experiment expects that there will be less growth in Caveolin than in Clathrin. Even so, both of the experiments use the same procedure of fluorescent microscopy.
Findings
The scattered dots observed in the first EGFR factor were more in number and larger in size than in the second experiment, with the endocytosis factor.

Task 5

Flow Cytometry
From the principles of cytometry, this experiment conducts the flow cytometry experiment with the labeling of Jurkat cells and the labeling of Phospho-Akt ser 473 antibodies. It uses three categories of cells, untreated, treated cells, and control experiments (Burgering and Coffer 1995, p54). All the cells are labeled in three different colors as they appear in the graph above. The red line is the control experiment, green is treated and blue is untreated. Read More