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Proteomics Seminar Series 2013 – Total Protein Loading Controls: A more reliable method for normalizing western blot data
September 18, 2013 @ 1:30 PM - 2:30 PM
“Total Protein Loading Controls: A more reliable method for normalizing western blot data”
Speaker: Patricia Piatti & Tom Davies, Product Managers, Bio-Rad
Presented by Bio-Rad and UF Interdisciplinary Center for Biotechnology Research.
Film has been the traditional method for the detection of chemiluminescent western blots. Its preference by researchers is based on mainly two features: good time-based sensitivity and resolution. Low initial cost has also contributed to its popularity, especially when many labs usually share equipment and consumables.
The development of digital imaging systems that use charged-coupled device (CCD) cameras to capture the luminescent signals has provided a new option for the detection and quantification of signals.
We will present data that shows digital imaging systems provide a superior method for capturing and analyzing chemiluminescent signals, with greater dynamic range, better limit of detection and more.
Loading controls are essential for reliable western blot data. Editors and reviewers of scientific journals are viewing quantitative western blot data with greater skepticism, often requesting raw data files to confirm that the proper controls have been used. This new requirement means that one has to either validate housekeeping proteins before using them as loading controls or find an alternative method that avoids this labor intensive and costly approach
This presentation will show:
- Why house-keeping proteins should be validated prior to use as loading controls.
- Examples of when house-keeping proteins are not appropriate loading controls and why.
- How total protein normalization can overcome this challenge by introducing a new and convenient technology to measure total protein load
Questions: Sixue Chen, Proteomics Division Director
Refreshments will be provided to attendees.