April 28, 2014
Associate Professor at the School of Life Sciences of Latrobe University
Western blotting has been used for protein analyses in a wide range of tissue samples for more than 30 years. Over the past few years, Dr. Robyn Murphy of Latrobe University has used skeletal muscle to develop a protocol for quantitatively analyzing proteins in very small samples sizes, here 1–3 mm segments of skeletal muscle fibers (individual muscle cells). This amounts to 5–10 µg total mass of tissue or 1–3 µg total protein. Prof. Murphy shows how this new protocol using Bio-Rad's stain-free gel technology ensures quantitative analysis in this unique procedure to examine fiber type-specific protein expression patterns.
Through the development of this technique, it has become apparent that using very small amounts of representative sample presents the most optimal situation for analyzing proteins. The introduction of Bio-Rad's stain-free gels has allowed the methodology to advance, whereby an assessment of total protein can be attained during the normal western blotting protocol. When combined with a calibration curve, quantitative assessment is achievable. In this webinar, Prof. Murphy demonstrates how western blotting can be reliable and quantitative and hence remain extremely valuable for making biological measurements.
About the speaker:
Robyn Murphy, Associate Professor at the School of Life Sciences of Latrobe University, leads a research group focusing on various aspects of skeletal muscle biochemistry. Her laboratory conducts studies in humans, using exercise and disease models, as well as animal models, with a focus on measuring proteins in segments of individual fibers to overcome issues with the heterogeneity of skeletal muscle.