As I had hoped, I will be working with hepatocytes today! In particular, through a combination of the techniques I previously learned, I will derive the protein concentrations of rat liver cytosol and rat liver homogenate in two samples and separate the proteins by molecular weight.
Part I: Protein Assay
These are the samples following incubation, from 0 – 2 microliters of BSA, and then sample 1 of rat liver cytosol to sample 2 of rat liver homogenate. As expected, the samples have changed from green to varying shades of purple through the reduction of Cu2+ to Cu+ by BCA and increasing concentrations of protein.
Part II: SDS Gel
Part III: Coomassie Blue Staining
This is the gel in the process of de-staining
And…this is the final product!
On the left is the rat liver homogenate and on the right is the rat liver cytosol. Wonderful! This de-stained more than the previous gel I ran, and the protein bands are more clearly visible. It’s always rewarding to see improvement!
I would like to give a special thanks to Peggy Wang for mentoring me throughout this process and Dr. Allan Wolkoff for providing the resources to work in his lab!