Melyssa Speeds Up the Core with Wes
"We’ve been able to increase productivity dramatically with Wes. The results are much better too, given that it removes the blotting step all together."
- Melyssa Bratton, Ph.D., Manager of the Cell, Molecular, and Biostatistics Core, Xavier University of Louisiana
Bridging the cancer incidence and mortality divide
Melyssa Bratton manages the Cell, Molecular, and Bioinformatics (CMB) Core facility, one of three core facilities in the RCMI Center for Cancer Research at Xavier University of Louisiana. A predominantly African American university, the RCMI program focuses on reducing disparities in cancer incidence and mortality between Caucasians and African Americans. Nationally, cancer rates are decreasing for Caucasians but increasing for African Americans who also have the highest rate of most major cancers across all racial and ethnic groups.
The CMB Core provides scientists at Xavier University with access to state of the art research equipment. They offer a wide variety of services including site-directed mutagenesis, bioassays, mammalian cell culture, and Western blot. The University’s vision is to achieve national prominence in the areas of cancer and health disparity research, and the core assists scientists in their research to eliminate health disparities among minority populations.
No way to get ahead with traditional Westerns
Melyssa’s group routinely used Western blots to test compounds for activity in a variety of cell systems from mammalian breast cancer cells to neuroblastoma cells. The core was normally working with at least 3–5 researchers needing Western blot data at any given time. That meant they were stretched thin running traditional Western blots as they didn’t have the time or the people to keep up with the number of targets they were asked to look at. Each blot needed a lot of hands-on time and took roughly two days to get results. And if the researcher also requested quantitation, the laborious data analysis process they had to go through made the process even longer. Melyssa’s group was spending all their time running traditional Westerns with no end in sight.
Boosting the core speed
Wes® gave the CMB core higher throughput and quicker turnarounds on results which let them dramatically increase their productivity. They then increased that even more by running samples from multiple projects together in one experiment. With Wes, each capillary pulls from its own source of sample, primary and secondary antibodies so it was easy for the group to probe for different targets in the same run. If they tried to do this with traditional Westerns, they’d have to cut the membrane into strips so each strip could be incubated with a different antibody.
Wes’ data was also more reproducible compared to blots since there was no need to transfer protein to the membrane either. And quantitation was super easy too using Compass for Simple Western.
Humming along at a new pace
The core is now able to process sample requests in a timely fashion. Melyssa’s group gets requests to study a wide variety of targets, so whenever they’re looking for a new target the Simple Western antibody database gives them a resource to find antibodies tested with Wes. Because ProteinSimple has three separation modules optimized to separate proteins within a broad molecular weight range (12–230, 2–40, and 66–440 kDa), she’s successfully looked at proteins ranging from 8 to 150 kDa to date (Figure 1).
The team’s also easily detecting several targets like phosphorylated Tau and phosphorylated EGFR with Wes that were hard to see on a traditional Western blot. They’ve only had the system for eight months, but have already published one paper1 and are working on another. All the free time Melyssa gets back with Wes lets her spend more of it on her other passions: reading and being a mom to an awesome kid.
Figure 1. Wes easily detects proteins from 8 to 185 kDa. (Left) EGF, an 8 kDa protein, was detected in a mutant cell line transfected with either wild-type or mutant endonuclease at different time points using the 2–40 kDa separation matrix. (Middle) Levels of ER alpha, a 65 kDa protein, decreased in MCF-7 cells treated with fulvestrant over the course of 24 hours (middle) using the 12–230 kDa separation matrix. (Right) HER2, a 185 kDa protein, was detected in cells transfected with a HER2/neu vector using the 66-440 kDa separation matrix.
A recent publication
1. J Liu, S Zheng, S Guo, C Zhang, Q Zhong, Q Zhang, P Ma, EV Skripnikova, MR Bratton, TE Wiese, G Wang, Rational design of a boron-modified triphenylethylene (GLL398) as an oral selective estrogen receptor downregulator, ACS Med Chem Letter, 2017: 8(1), 102-106.