#5. Keep It Clean
Your Western blot results can be greatly affected by your attention to detail. Make sure that your incubation boxes and forceps are thoroughly cleaned, rinsed, and dried before you start. Handle membranes only with forceps to prevent fingerprints that are easily seen with fluorescent detection. Do not pour your reagents directly on your blots; instead, pour down the sides of the incubation box or container. Write on blots with pencil, not ink, to prevent unwanted smears and spots. When imaging, be sure your imaging surface is clean and lint free to prevent speckling, smearing, and contamination by other dyes (e.g., Coomassie blue, fluorescent dyes, or residual substrates). Prevent introducing background with chemiluminescence detection by wicking off excess substrate to prevent pooling on the membrane.