I haven't been in the darkroom for months. It is nice not to have to wait in the hallway while someone else is using the darkroom.
Dr. Lisa Keyes
University of Florida College of Medicine
At the University of Florida’s College of Medicine, Dr. Lisa Keyes is exploring molecular channels of host-pathogen interactions. As a postdoctoral fellow in Dr. Scott Tibbetts' lab, Keyes is decrypting the interaction between viral non-coding RNAs and epigenetic machinery in a murine herpesvirus model.
Murine gammaherpesvirus 68 (gammaHV 68, genetically associated with human herpesviruses) causes lymphoma in mice, and makes for a useful model system to study host-pathogen interactions. Keyes is specifically investigating the mechanism through which gammaHV 68 exploits the host epigenetic machinery during pathogenesis. “Viral long non-coding RNAs interact with histone deacetylases and other epigenetic proteins in order to induce replication or latency depending on the cell type1. These non-coding RNAs may act as scaffolds for host proteins,” Keyes says.
In looking at the expression and interaction of a variety of viral and host proteins, Keyes performs as many as 30 Western blots each week. Their lab houses a C‑DiGit® Blot Scanner for visualizing all of their Western blots.
Valuable Solutions for Chemiluminescent Western Blotting
Tibbetts' lab learned about the C‑DiGit Scanner during a LI-COR seminar at the University of Florida campus. As luck would have it, they won their instrument in a giveaway. Keyes has the opportunity to use the instrument extensively and finds it valuable. “The C‑DiGit Scanner has saved tons of film and money, and more importantly our time in trying to get the right exposure. It has really generated cleaner and easy-to-read results.”
The C-DiGit Scanner has saved tons of film and money, and more importantly our time in trying to get the right exposure. It has really generated cleaner and easy-to-read results.
The instrument is popular not only among other lab members but also with neighboring labs. “Others in our lab and especially first-timers, who have never done blots before, like it because it is streamlined and straightforward. We work closely with the lab next door and they have used it quite a few times. They really like it too,” she says.
The lab uses WesternSure® PREMIUM Chemiluminescent Substrate for detection, and the WesternSure® Pen comes in handy to annotate and label blots. “It is so nice to be able to just draw right on the membrane and then see it [when scanning]. I never have to worry about lining up [the blot with the ladder reference] as I know where the ladder lies in reference to the sample. I can mark the membranes and keep track of the numerous blots that I run every week,” says Keyes.
Reliable Digital Imaging of Western Blots
Keyes is happy to have moved away from developing blots on film. “On film, the blot might be a little bit darker one day and the next day a little lighter. If you are trying to re-expose it, you never really get the same blot over and over again. I have noticed with the C‑DiGit Scanner that you can redo it and everything else still looks the same when you put it back on, so it's pretty nice.”
To Keyes, data from the C‑DiGit Scanner conveys reliability. “It gives me more confidence. I know what I am looking at isn't an artifact of just how something went through in the machine.”
To top it all off, Keyes doesn’t need to make trips to the darkroom anymore. “I haven’t been in the darkroom for months. It is nice not to have to wait in the hallway while someone else is using the darkroom.”
Researchers like Keyes are obtaining reliable data with user-friendly C‑DiGit Blot Scanner technology.
1Feldman ER, Kara M, Coleman CB, Grau KR, Oko LM, Krueger BJ, Renne R, van Dyk LF, Tibbetts SA (2014). Virus-encoded microRNAs facilitate gammaherpesvirus latency and pathogenesis in vivo. MBio. 2014 May 27;5(3) :e00981-14. doi: 10.1128/mBio.00981-14.
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