Investigating Resolution of Inflammation with Jess
"Jess is a user-friendly, multi-purpose platform that gives me the possibility to perform Western blot analysis on precious samples, as well as quantifying proteins expressed in the cells at the same time." Searching for key inflammation molecules"
- Kirstine Nolling Jensen, PhD Student, Department of Immunology, Landspitali, the National University Hospital Iceland
Searching for key inflammation molecules
Chronic inflammatory diseases, such as diabetes and asthma, are leading causes of death around the world. Understanding the biological mechanisms and cell signaling pathways involved in inflammation holds the key to improving treatments and saving lives. One phase of the inflammatory response that remains poorly understood is the resolution of inflammation. Only in recent years has it come to light that this process is actively controlled by interactions between innate immune cells, such as natural killer cells and neutrophils, and other non-immune cells. Through conducting further studies into the interactions of these immune cells in the termination of inflammation and the restoration of tissue homeostasis and functionality, the process of resolving inflammation can be more thoroughly defined.
Kirstine Nolling Jensen is a PhD student at the Department of Immunology at Landspitali - the National University Hospital Iceland. She is currently investigating the role of natural killer (NK) cells and neutrophils in the resolution of inflammation, as well as spending time monitoring the immunological status of patients, as part of the hospital’s flow cytometry service.
Through her research, Kirstine is seeking to identify key molecules expressed by natural killer cells and neutrophils that contribute to resolution of inflammation. In particular, she is looking into signaling pathways in these cells in response to omega-3 polyunsaturated fatty acids consumed through diet or added to culture before active inflammatory responses and resolution.
Running into quantitation difficulties
Due to the precious nature of her samples, Kirstine often struggled to get clear results that could be quantitated with traditional Western blotting, as the resulting bands were very weak. This prompted Kirstine to find a new way to perform these assays, something where she could use smaller samples and gain reproducible results with less hands-on time.
Small samples go a long way
By running automated Simple Western™ assays with Jess, Kirstine can use as little as 3 μL of each sample and yet still achieve the high signal:noise ratios she requires. After completion of each assay, Kirstine is then able to confirm protein expression in her cells using RNA sequencing datasets and LC MS/MS.
With Jess' built-in analysis software, protein quantitation is a breeze. Kirstine has gone from struggling to get results after 2 days of manually carrying out Western blots to receiving fully analyzed results on 24 samples in just 3 hours with Jess. This lightning speed turnaround time combined with only 30 minutes of hands-on experimental time for each Simple Western assay means Kirstine has freed up her time to spend more time on interpreting results, with the hope of publishing her next paper soon!
The user-friendly platform of Jess means that once assays are set up, it is simple to reproduce and quantitate results. If Kirstine did ever run into issues, the ProteinSimple team are always on-hand to assist with troubleshooting.
A brighter future for immunoassays?
By utilizing Jess as her protein analysis solution, Kirstine has not only reduced her use of single use plastic by using fewer pipette tips, but also stopped using toxic chemicals associated with traditional Western blotting, making it a much greener solution.
Since using Jess, Kirstine has helped two other lab groups to get started with running Simple Western assays in their research and looks forward to bringing more colleagues into the world of automated Westerns, making everyone’s lives that little bit simpler.
With the acceleration of these inflammation research projects at Landspitali, Kirstine and her colleagues are outputting more data and analysis at a faster rate. These studies may pose answers to many of the clinical questions surrounding inflammation which could, in time, lead to better treatments and a stronger understanding of diabetes, asthma, and other chronic inflammatory diseases.
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