Western Blot eHandbook
View our step-by-step guide to western blotting including troubleshooting tips.
Step-by-Step Guide to Western Blotting
Understand the principles behind this common technique and learn how to achieve reliable and reproducible results.
Whether you are a student/researcher new to western blotting, or a skilled scientist, you’ll gain valuable insights to help you to master the technique.
- Main principles of western blotting
- Troubleshooting tips
- How to accurately analyze data
- Proper controls, antibody validation, and data normalization
- Fully automated Simple Western™ platforms from ProteinSimple, a Bio-Techne brand
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Problem: High Uniform Background
Possible Reason & Solutions: Non-specific binding due to high antibody concentration
- Lower concentration of primary or secondary antibody (titrations may be helpful).
- Use a strong blocking agent in antibody diluents (milk is typically a stronger blocking agent than BSA).
- Confirm that the background is not due to issues with the secondary antibody by omitting the primary antibody.