Hey everyone! For the past few weeks, I have completed a variety of tasks and assays that makes each day in the lab unique. So, for this week’s blog, I’ve chosen to reflect on what I did and what I experienced in the lab last Friday as it was both an eventful and enjoyable day!
During most of last week, I was completing a protein purification, an important multi-day process that allows me to obtain pure, but small, amounts of protein that I can subsequently test. The protein of interest is βarr1-393, a truncated version of wild-type βarr1 at the 393rd amino acid residue. Most of this work was done with the help and assistance of my good lab friend Brittany, whom I would also like to thank for showing me how to understand and complete this important process!
Luckily for both of us, the most tedious and time-consuming part of the purification happened on Thursday. I arrive at the lab at around 10 AM and the first thing we need to do is dilute our eluate (currently at 300mM) from the previous day in preparation for ion-exchange chromatography. I grab four 30 kDa filters and equilibrate them with wash buffer. Then, I add the protein eluate from the day before, spin the filters down, and add dilution buffer in a slow, drop-wise fashion until we have our desired 50mM concentration.
This final step (dilution via a drop-wise fashion) is when the part of the waiting game begins. Luckily, this is the ideal time for me to grab lunch and chat with Brittany about her week and mine as well. These conversations are always nice to have as it allows me to take a quick break from lab and make personal connections with my colleagues!
After dilution is complete at around 1 PM, we load our sample into the ion-exchange chromatography machine. A lot of the work here is done on Brittany’s part, and I still have a lot to learn regarding how to use the machine in other contexts. This process generally takes 2-3 hours in total, giving us more downtime during the day to complete other things we might have planned.
Our Fridays end with a weekly lab tradition called Porters. Dr. Lefkowitz comes around each room with a little bell signaling that it’s time to meet. Most of the lab members come into his office, sit down, and chat about anything that has been on our minds for the week. For this week specifically, however, one of Dr. Lefkowitz’s closet colleagues, Dr. Howard Rockman, joined us. As a renowned cardiologist, Dr. Rockman brought us copies of an ECG of a recent patient that he examined and he gave us a quick lesson on how to analyze the chart and diagnose the patient. It was such an intriguing conversation (as I am in LOVE with the cardiovascular system) and I am so thankful that I was able to experience such a time.
The end of my day ended with me talking with my mentor Bullet, who talked about what our plans were for the upcoming week and what I needed to prepare and/or read during the weekend. As I walked out of lab, I reflected on my week as a successful one and anticipated the new things I will learn the next time I enter the doors of the CARL building once again!
