Detecting Low Abundance Proteins in Precious Pediatric Cancer Samples with Wes
"Some of the proteins we wanted to detect are low in abundance and it’s really hard to get a good amount of protein for traditional Western blot. The low protein concentrations required by Wes makes it easier to save precious samples ensuring protein detection."
- Debbie Hicks, Ph.D., Faculty Fellow, Wolfson Childhood Cancer Research Centre, Northern Institute for Cancer Research, Newcastle University
Advancing the treatment of childhood cancers
Work of the Paediatric Brain Tumour group (PBTG) at the Wolfson Childhood Cancer Research Centre focuses on developing highly effective treatments for childhood cancers, reducing both toxicity and side effects of treatments. Debbie Hicks, Faculty Fellow, studies the molecular basis of pediatric brain tumor development and the application of molecular diagnostics for improved individualized treatment regimes. Together with graduate students Azira Ramli and Gemma Llargues-Sistac, they aim to develop novel therapies for the treatment of pediatric tumors.
High-quality protein detection from precious samples
The PBTG used traditional Western blots to analyze their proteins of interest but struggled with protein detection in their limited samples. They implemented Wes™ to detect their proteins of interest from nuclear, cytoplasmic or total protein extractions faster and more efficiently than they could with traditional Western blotting. The low protein concentrations required by Wes made it easier detect proteins of interest in their precious samples. They plan to expand the use of Wes to validate the expression levels of key proteins in their pediatric brain tumor cell line models.
One of the proteins under investigation is very small, 16 kDa, and hardly detectable by conventional Western blotting. Wes offers a higher resolution that enables the detection of this protein using as little as 0.8 μg/mL of cell lysate.
Saving sample, time and money
When working with traditional Western blots, it took the team three days to generate results, and sometimes the experiment needed to be repeated as they were unable to detect any protein on the membrane. With Wes, they can run up to 25 samples at the same time, using very low concentrations of lysate. Moreover, with multiplexing, they can measure multiple proteins at the same time, saving time and enabling them to get the most out of their samples. The team gets quantitative results in just three hours, without all the hands-on steps associated with traditional Westerns. The in-built quantification system enables them to accurately detect, quantify and analyze results without the need for additional software. In addition to the sample and time savings, the researchers noticed they were saving money on antibody costs due to a reduction in the amount of antibody required to detect their proteins of interest compared to the amounts needed in traditional Westerns.
Wes is keeping busy at Wolfson Childhood Cancer Research Centre. Based on the impressive results from the PBTG, the technology is being implemented by many different research groups at the Centre.