Individual skill
western-blot-troubleshooting
Western blot optimization and troubleshooting — antibodies, transfer, detection, and quantification done right.
skills/western-blot-troubleshooting/SKILL.md
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- License
- MIT
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## Overview
western-blot-troubleshooting covers getting reliable western blots: gel and transfer
optimization, antibody validation, detection methods, and the quantification discipline that
separates data from pretty pictures. Western blotting is semi-quantitative at best — this skill
is about staying within the "quantitative" part honestly.
## When to use
- Optimizing new antibodies: dilution, incubation, blocking.
- Transfer problems: incomplete transfer, blow-through, uneven transfer.
- Detection issues: no signal, high background, non-specific bands, speckles.
- Quantification: linear range, normalization (total protein vs housekeeping), densitometry.
- Antibody validation: knockout/knockdown controls, blocking peptides.