Trustee Presentation

On Friday (February 21st), I was again unable to attend my internship because it was the day of the Emma Talks! Emma talks offered a very unique opportunity to hear from six of the most impactful women in the world speaking on topics from the environment, to women in the workplace, to the importance of girls' education.

After the Emma Talks, I had the opportunity to present my own work from my internship to the Board of Trustees! It was a great opportunity to share what we're working on.

I look forward to getting back into the lab this week and starting my blocking buffer experiment!

Snow Day!

On Friday (February 14th), I was unable to attend my internship because we had a snow day! Instead of any scientific updates, here's a beautiful picture of Emma in snow!

Testing Dye Binding

Yesterday (February 7th), I continued work on my new project. I ran a preliminary test to confirm the way in which dye binds to slides with PBS and Casein. From this experiment, we expected that the slide incubated with casein would be stained, while the slide incubated with PBS would not. To test this, I incubated one slide with 3 mL of 10X Casein and one slide with 3 mL of PBS for an hour on the shaker. Once the incubation was complete, I drained the solutions into the biologic waste container and washed both of the slides with PBS three times for 10 minutes each time. I then added 3 mL of LabSafe GEL Blue stain onto each slide for 20 minutes.

Once this process was complete, we found a surprising result. The PBS slide, as expected, had not bound the dye. However, the Casein slide did not bind the dye either. Instead, the protein had bound the dye, and the protein was no longer bound to the slide. In the image below, the Casein slide (left) and PBS slide (right) are both pictured with the dye solution still on the slides.

After seeing this result, we decided that either a small amount of Casein was stuck under the slide and came up to bind the dye, preventing us from seeing the protein on the surface of the slide, or the dye eluted the Casein from the slide. In an attempt to see if there was still protein bound to the surface of the slide, I poured the dye off of the slide, washed it with PBS, sprayed it with water, and added 3 mL of dye back onto the slide in a different container. After 10 minutes, the slide showed no change in color, leading us to believe that there was no protein bound to the slide. We will test another Casein slide to see if we get the same result. If the dye is found to elute the Casein from the slide, we will need to reconsider the procedure for my project.

While waiting for the slides in this experiment to incubate, I labeled the trays for my project and prepared my slides by washing them in ethanol.

I look forward to finding out if the dye does indeed elute the Casein from the slides.

First Step: Blocking Buffer Solutions

On Friday (January 31st), I worked on the first steps of my new blocking buffer project! Because I did not specifically explain the purpose of a blocking buffer in my previous posts, blocking buffers are used to prevent nonspecific binding, reduce background signal, and stabilize proteins for better interactions. These effects are demonstrated in the following image.

Image from http://www.immunochemistry.com/products/elisa-solutions-1/blocking-buffers.html

Today I took my first step in my project by making my blocking buffer solutions! We decided to make 5 mL of each BSA solution (5%, 2.5%, 1%, 0.2%) and 10 mL of each Casein solution (1x and 1/2x). First, I had to do the calculations to determine the amounts of the buffers and the amounts of PBS that I would need for the dilutions. First, I would make the highest concentration solution and dilute from that. Below are my calculations.

Once the calculations were confirmed, I completed the dilutions. I have now officially started my own project! I can't wait to carry out this experiment!