Altered phosphorylation levels in signaling pathways regulating various cellular functions — such as cell growth, differentiation, and apoptosis — are strongly associated with cancer. Detecting protein phosphorylation events and aberrant phosphorylation levels is therefore essential for research into various biological and pathological processes.
However, detection of phosphorylated proteins by western blotting can be challenging. In this 45-minute webinar, you will learn the important steps to obtain clean western blots of phospho-proteins. These include:
- Selecting antibodies
- Preparing lysate samples
- Blotting and washing buffer selection
- Applying best practices for phosphatase treatment
- Determining cross-reactivity to total proteins
- Choosing the best controls
- Quantifying phosphorylation levels using total protein normalization
Rachael Preston, PhD,
Applications Scientist at Bio‑Rad Laboratories
Rachael earned her PhD in cancer genetics at Cardiff University (UK) where she focused on characterizing the tumor suppression function of folliculin. Of particular focus was the detection of phosphorylation events resulting from the activation of kinases, like mTOR and AMPK. She has extensive experience in molecular biology techniques and antibody applications.