Throughout the past three months, I have been investigating a process in which a certain human protein can be easily detected through an efficient, inexpensive ELISA protocol. This protein, PKR, is crucial to the human immune system and is often correlated to the proliferation of certain fatal diseases. My contribution to this field includes constructing a protocol in which this specific protein can easily be detected in a variety of human cells. The standard ELISA procedure is very simple and easily interpreted if the proper conditions are administered. The ELISA is dependent upon a sandwich combination of antigen, antibody and an enzyme substrate that is deposted in a 96 well microtiter plate.
After repeated trials of a standard ELISA protocol, I manipulated several variables to promote the upregulation and detection of the specific protein. One of the crucial components of my success with the ELISA has been interferon. Interferon (IFN) is known to induce the protein kinase PKR and promote expression severfolds. When comparing the non-treated cells vs. treated cells, there is a significant difference in the expression of PKR. IFN has allowed us to discover a significant presence of PKR in daudi cells, which can potentially lead towards producing an ELISA that can easily detect the specific protein.