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Line 30: == Methods == The dot blot was originally performed on a piece of [[nitrocellulose]] membrane or PVDF membrane. After the protein samples are spotted onto the membrane, the membrane is placed in a plastic container and sequentially incubated in blocking buffer, antibody ~~solutions~~solution, or rinsing buffer on a shaker. Finally, for [[chemiluminescence]] imaging, the piece of membrane is wrapped in a transparent plastic film filled with enzyme substrate. A vacuum-based dot blot apparatus (Bio-dot or Bio-slot from BioRad) has been used to facilitate the rinsing and incubating process by using vacuum to extract the solution from underneath the membrane, which is assembled in between several layers of plates to ensure good seal between sample wells, hold waste solution, and deliver suction force.{{citation needed\|date=November 2017}} For chemiluminescence signal detection, the apparatus is dissembled and the membrane is taken out and wrapped in a transparent plastic film filled with enzyme substrate.

Dot blot: Difference between revisions