Designing a Quantitative Western Blot Experiment to Avoid Common Pitfalls

Can We Trust Western Blots?

Using Bio-Rad’s PDQuest™ Software to Generate a Match Rate between 2-D Electrophoresis and Western Blotting

Two-dimensional (2-D) electrophoresis is a widely used gel-based method for separating protein mixtures by charge (isoelectric point) and size (molecular weight). After 2-D electrophoresis (2-DE; described in detail in the Bio-Rad Applications & Technologies section), gels can be processed for protein detection via total protein staining and/or western blotting. Western blotting analysis following 2-DE separation is particularly useful in at least two important applications: (1) identification of pathogen-derived proteins that are highly immunogenic (have potential as vaccine candidates), and (2) evaluation of the range of host cell protein (HCP) impurities detectable by polyclonal antibodies used to monitor HCP impurities in purified biopharmaceuticals. For both applications, it is critical to generate a match rate from an overlay of the 2-DE and western blotting analyses. Bio-Rad’s PDQuest 2-D analysis software can be used to easily generate match rates. We describe here, step-by-step guidelines for generating this information using PDQuest software.

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Summer Student Tutorial: How to Avoid a Leaky Gel

This week’s summer student tutorial is something that we have all experienced in the past. Leaky gels. While the best solution is probably to purchase a new pad, for the more resourceful scientists out there, here is a tip for you.

The video is courtesy of LabTricks which has no affiliation to Bio-Rad Laboratories.

Summer Student Tutorial: Running a Mini-PROTEAN Gel

Welcome to the third installment of the summer student tutorial series where summer students learn essential lab techniques without sapping the resources of their graduate student lab-mates. In this edition, you will learn how to set up and run a mini-PROTEAN TGX Precast Gel. Enjoy! And once again, if there are other tutorials you would be interested seeing in this space, please let us know and we will try to get it online as soon as possible.