Western Blotting
Western blotting is a well-established technique for detecting, analyzing, and quantifying proteins. However, Western blots often suffer from poor consistency, high background/low sensitivity, and lack of robustness. While blotting should be a science, it can seem like an unconquerable art. For streamlined Western blotting with high signal-to-noise ratios, think about the principles behind each step:
- Sample Preparation: use optimized extraction reagents and protease inhibitors; accurately quantify total protein
- Electrophoresis: obtain clear and sharp blots by using high-resolution gels
- Transfer: use the correct transfer membrane that’s best for your analysis and detection method
- Immunodetection: optimize blocking and washing; use Western-tested antibodies and detection substrates for the highest signal/background.
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