This past week at my fellowship, I completed immunofluorescence staining by myself. Although some biochemical experiments may be easy enough to follow the protocol, the samples I was working with required attention to detail and being careful while making the proper ratios of reagents.
Since our staining last week showed too much background, we began troubleshooting the experiment. We had four groups each with a different set of conditions. The samples were fixed in methanol or acetone and treated with TritonX-100 or without TritonX-100. After microscopy, I was able to identify two treatments that maximized more specific staining for one of our proteins of interest (protein #1). The second protein of interest was only seen using one of the treatments. Oddly enough, it was one of the treatments that worked for protein #1. This makes me wonder if these conditions are the most optimal for other antibodies in which we have worked.
I have spent the last few Mondays presenting in lab meetings so I have been trying to follow the protocols exactly to get good quality data to present. I am excited to start working on my last presentation which is supposed to be a ten minute presentation about what I studied this summer.