Characterization of Auto-Antigens as Biomarkers in Kidney Cancer

Provide a 19 pages analysis while answering the following question: Characterization of Auto-Antigens as Biomarkers in Kidney Cancer. Prepare this assignment according to the guidelines found in the APA Style Guide. An abstract is required. Furthermore, the serological Expression Cloning i.e. SEREX is used to diagnose a tumor in humans causing elevated IgG antibodies.

In the clear cell kidney cancer cell Caki-1, TROVE2 protein is described in isoform. In order to start this experiment Caki-1, cells are cultured under restricted conditions such as typical growth and medium standard on average supplemented 0.02% DMSO for 72 hours. Now after 24 hours, 48 hours, and 72 hours the protein is extracted while performing western blot by using monoclonal antibody elevation alongside N-terminal peptide of the human TROVE2 protein.

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Well 3 1.06×10^5 cells/mL – 2.01×10^4 cells/mL 20% viability. Not an obvious phenotype to indicate this level of cell death. Transfection may have work, but not sure. No protein samples prepared through Phenotypes.

In the phenotypes, the difference is not observed after passing 24 hours and 48 hours. In well 3 the positive control is seen after passing 48 hours. But this is present less than the controls and the difference is calculated as 20%. In order to release each well, 1ml of TrypPLE is used at 48 hours. After completing this process 1ml of media is added and 2ml cells are removed from the tubes. The cell count is done after finishing all the steps.

The gel is run at 25μg of well 2, 3, 4, 5, and 6 RIPA is extracted at 4-20% TGX gel 300v. The TBT is transferred by using a brand new machine and mixed weight settings are used. Probed overnight along with BlotCycler recycling 12.5mL of clarified antibody aliquot with added anti-Trove2 (mouse monoclonal ab58004 1/2000) and anti-SSB (Rabbit monoclonal ab124932, 1/2000). Secondary’s ab96882 and ab96884 Antibodies and Antisera.

The growth media named McCoy’s 5a that is altered on an average basis and enhanced along with fetal&nbsp.bovine serum (FBS). The FBS is concentrated to 10% and x1 Gibco X100 Antibiotic-Antimycotic solution is added.

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