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Western Blot / WB tips
- 1. General Tips to consider when performing a western blot
- 2. Sinobiological Western Blot Conditions
- 3. Why is the actual western blot band size different from the predicted?
- 4. Why do I see a weak signal or no signal at all on my western blot?
- 5. Will this antibody react with the same target in a different species?
- 6. Why are there extra bands on my western blot?
- 7. Do my transfer conditions change if my protein of interest if >180kD?
- 1. Determine the best usage of the target protein and primary antibody
- 2. Keep up the protein transfer efficiency
- 3. Anticipate the effect of gel thickness in western blot
- 4. Make sure to equilibrate membranes and gels on transfer solution
- 5. Cleaner blots with the right blocking solution
- 6. Optimize your incubation time
- 7. Sometimes antibodies won't recognize denatured proteins
- 8. Western blot of phosphorylated proteins
- 9. Detection of low abundance proteins
- 10. Stripping and re-probing membranes