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Line 12: == Methods == A general dot blot protocol involves spotting ~~1-2~~1–2 microliters of a samples onto a [[nitrocellulose]] or [[PVDF]] membrane and letting it air dry. Samples can be in the form of tissue culture supernatants, blood serum, cell extracts, or other preparations.<ref name="abcam">{{cite web\|url=http://www.abcam.com/ps/pdf/protocols/dot%20blot%20protocol.pdf\|title=Dot Blot Protocol}}</ref> The membrane is incubated in blocking buffer to prevent non-specific binding. It is then incubated with a primary antibody followed by a detection antibody or a primary antibody conjugated to a detection molecule (commonly [[horseradish peroxidase\|HRP]] or [[alkaline phosphatase]]). After antibody binding, the membrane is incubated with a [[chemiluminescent]] substrate and imaged.

Dot blot: Difference between revisions