Group C: Levels of Endogenous Cofilin

Description: 

In this experiment, we want to want to compare the levels of endogenous cofilin and TEVp-Cofilin expressed under an inducible promoter with different levels of dox induction. This will allow us to determine what level of expression of cofilin would lead to the best signal:noise ratio of exogenous cofilin to endogenous cofilin. We are planning to transfect HEK293 with TEVp-Cofilin  under a TRE promoter and induce with varying levels of dox. We will then preform a Western blot with an anti-cofilin primary antibody and compare band sizes to test for the relative amounts of endogenous cofilin and exogenous TEVp-Cofilin  in the cells.

Transfection Plan:

Stuff:

GAPDH: Loading control (how many cells we've loaded onto the gel)

GFP: Transfection marker (how many transfected cells we've loaded onto the gel)

We are planning to get the 4-25% gel in order to optimize our resolution.

Expected Results:

http://www.thermoscientificbio.com/uploadedFiles/Resources/general-recommendations-for-sds-page.pdf