Covers Many Western Blot Problems and Includes Many Solutions. Fuzzy Bands, Low or Weak Signal, High Background, Spots on Film, Too Many Bands. A MolecularStation Guide. - [Read Troubleshooting Western Blot]
For cells grown in tissue culture, the most useful method of lysis is treating with detergents, as described in this protocol. Non-ionic detergents, such as NP-40, solubilize the plasma and intracellular membranes, break many weak intermolecular bonds, and solubilize most of the commonly studied protein antigens. RIPA lysis buffer may be used as a more rigorous extraction buffer to release all but the insoluble proteins of the cell and to break most weak noncovalent interactions. - [Read Lysing Tissue-Culture Cells for Immunoprecipitation Protocol]
Troubleshooting Guide for Chemiluminescent Western Blotting Pierce. High Background, Reverse image on film, Blot glows in the darkroom, Signal duration is less than 8 hours, Weak or no signal, Spots within the protein bands, non-specific bands. - [Read Troubleshooting Guide for Chemiluminescent Western Blotting Pierce]
Streaking of Western Blots, Weak Bands or Weak Staining of Western Blots, Poor Transfer of Membranes, Dirty Blot, Multiple Bands on Western Blot Membrane and Film. Western Blot Info. - [Read Western Blot Troubleshooting]