The project this week involved using a modified Lowry Protein Assay method for protein determination. The results were to be examined by both the traditional spectrophotometer and an Iphone camera. On tuesday the results were not accurate. Our standard solutions did not produce the necessary color after the Folin reagent was added. We were confused and spent the rest of the day backtracking our experiment to see what might have gone wrong. We looked over all our calculations and solutions added and found we added too little BSA protein. We adjusted and ran the standards but still there was no color indicating protein was present. Brenna and I decided to call it a day and Aj. Gate's lab group told us they would try to find an explanation.
The next day Pi Oum figured out that the procedure used had a typo. For our alkaline solution to add into the standards, we were preparing it with 1M NaOH instead of 0.1M NaOH. This major difference in concentration had to be the reason our solutions weren't changing color. We applied the changes and turns out our standards worked! The solutions displayed color and the absorbance values were linear with concentration. All was good until we next ran the unknown solutions. When things were running smoothly again we hit a wall. We became frustrated and ended up staying till 7pm with our lab group to figure out what possibly went wrong.
After dinner, Brenna and I decided to go to a cafe to discuss our experiment. We ran into Dr. Sandquist and he offered his advice on how the unknown solutions of protein may have deteriorated over time sitting in the lab. Brenna read from some articles about the Lowry method and read the importance of vortexing the solutions immediately after mixing and the incubation time for each sample. We then emailed Oum our plans for the experiment in hopes of getting it correct the next day.
The next day we prepared all the standards and received freshly made unknowns to measure. With my bad luck I got sick and was unable to finish the 2nd half of the experiment. However, Brenna finished the 2nd half promptly and efficiently. We calculated our results from the spectrophotometer and turns out we calculated the correct concentrations for our unknowns! All our struggles came to be worth it as it turns out we were successful!
Also to brighten my day my labmates bought me medicine of a thai milk of magnesia with menthol. It tasted kind of funny, but the results of the medicine made my pains go away. By night I was able to walk the local night markets freely without any stomach aches. I Don't exactly know what was inside the medicine, but I'm glad of the results.
After the long week of ups and downs I’m just glad it’s the weekend!
Still hanging on!