Western Blot Analysis with Rat Liver Homogenate

Western blot analyses are run to separate and identify proteins. First, the proteins are separated through gel electrophoresis. The proteins are transferred to a membrane where they are marked with antibody, and a film is developed for visualization (‘Western Blot: Technique’). Gel preparation: Waiting for the gel to solidify: Adding the samples of Rat liver homogenate into the gel: Waiting for the gel to run: At the beginning: Almost done: Here, the protein is being pushed through the gel by an electric field. The purple dye is used to identify the edges. The next step is blotting, where the proteins … Continue reading Western Blot Analysis with Rat Liver Homogenate

Cell Culture Maintenance

Cell cultures must be fed every few days with fresh growth medium, and that which they have already consumed must be disposed of. This is the HeLa cell culture I created last week. The solution has changed from pink to yellow, as the cells have consumed the growth medium. On the left is the growth medium, in the middle is PBS, and on the right is Tripsin. I dumped the old medium into a beaker, then washed the plate with PBS and tripsin. Next, I detached the cells from the plate and added them to a new plate with fresh … Continue reading Cell Culture Maintenance

Coomassie Blue of Rat Liver Cytosol and Homogenate

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 … Continue reading Coomassie Blue of Rat Liver Cytosol and Homogenate

HeLa Cell Cultures!

These are HeLa cell cultures. (Mine is on the left) And here are more, being frozen at -80°C HeLa cells were derived in 1951 from the cervical cells of Henrietta Lacks, a terminal cancer patient. Scientist George Gey extracted and cultured the cells in a manner which allowed them to live indefinitely. For the past six decades, HeLa cells have been used in medical research for purposes primarily relating to curing diseases (‘Henrietta Lacks’). We can thank HeLa cells, and those who contributed to their creation, for numerous scientific breakthroughs throughout the world, including those achieved in this very lab! … Continue reading HeLa Cell Cultures!

Research at the Albert Einstein College of Medicine

I have recently begun volunteering at the Albert Einstein College of Medicine Department of Gastroenterology and Liver Related Diseases. The department is run by Dr. Allan W Wolkoff, whose lab I am fortunate enough to work in. His research concerns organic ion uptake by hepatocytes. During my time here, I will learn basic research techniques and conduct an independent research project. I began by learning to use a pipette. On my first day I transferred coomassie blue dye into microcentrifuge tubes in exponentially increasing volumes of 2 microliters from 2 to 512. After being able to accurately insert the required volumes … Continue reading Research at the Albert Einstein College of Medicine