Your Professor has asked you to design an experiment to determine if a particula

Question

Your Professor has asked you to design an experiment to determine if a particular protein (CSI 1) is present in both Zebra fish and Gold fish brain. Using one page describe how would you design an experiment to identify, locate and characterize (CSI 1). Since the particular protein CSI 1 is found in the both Zebra fish and Gold fish brain there can be two approaches for this experiment. However the best way to approach this is by identifying the particular protein by Immunohistochemistry, this process will not only identify the protein but will also located it with the fluorescence tag. The process will include tissue preparation + fixation, antibody addition (monoclonal since we are looking for a particular protein), add a fluorescence tag to the antibody and we can locate the interaction because of color. We will begin by taking brain tissue from both of the fish and put them in separate plates. Anti- CSI 1 is added to both plates so that we can determine whether the proteins are present (positive control) or not present (negative control). The Anti-CSI 1 is fluorescence tagged. This will create an Anti-CSI 1-CSI 1 Protein complex an Antibody/Antigen interaction. This can help locate where this protein is present in the brain cell because it will fluorescent. Once the protein is localized we can extract it by protein purification. The extracted protein is purified and can be characterized through Mass spectroscopy, which is the fastest way in determining protein sequence. Once a protein sequence is known then its characterization can be predicted. When using Mass Spectroscopy, the entire primary sequence of the amino acid can be known because it gives us information on its interaction and folding based on posttranslational modification.

Explanation / Answer

The presence of proteins can be determined by flourescent immuno histochemistry. The precise location of proteins can be determined by flourescent labelled antibodies which will cause antigen antibody reaction. After determining the location, proteins can be purified and characterised by mass spectroscopy.

Therefore, the experiment described in the question is correct.

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