Today we started a new western blott with the samples we prepared yesterday. This morning, we first prepared the samples by adding ethanol to rehydrate the peptides and sample buffer. We then vortexed them to resuspend the proteins, poked holes in the top so that pressure would not build up in the tubes during heating and then put them in the incubator at 55 degrees Celsius. After the samples were prepared we put them on ice and loaded them onto the gel to be electrophoresed. We ran the current through the gel for about three and a half hours. After that, we prepared the western blott sandwich by cutting the blotting and nitrocellulose paper and soaking them in 10% methanol. Then we removed the gel and put the sponges, blotting paper, nitrocellulose paper, and gel into the sandwich. The proteins are being transferred from the gel to the paper now and will be ready around 4:05. After the paper is ready, we will remove it from the sandwich and set it out on our lab space to dry overnight.
Dr. Brown gave me some extra textbooks today on endocrinology that should help with the primary literature I have been reading. I have been reading one of the books during some of the down time today.
I also spoke to my dad last night and he said that he had arranged for me to shadow Dr. Harper, general surgeon on Friday.
Tomorrow we will label the paper and do the primary and secondary washes over our western blott.
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