Single-Cell Western Blot and Stain-Free Total Protein Loading Control



February 23, 2017

"For more info, visit Robyn Murphy, Associate Professor at the School of Life Sciences of Latrobe University, has developed a unique protocol to conduct western blotting on muscles at single-cell levels to examine fiber type-specific protein expression patterns. Find out how Bio-Rad’s stain-free gel technology helps ensure quantitative analysis in this unique procedure. 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, Prof. Murphy 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. 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."

Analytical Techniques

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