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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
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