Immunofluorescence Counterstain - PI Counterstain Solution

Tony Rook

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Nov 16, 2015
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Please find the following media preparation method:

Immunofluorescence Counterstain:

PI Counterstain Solution

PI Stock Solution (1mg/ml or 1.5 mM):
To prepare, add 1 mg PI (Propidium Iodide) to 1 ml distilled water. Store stock solution at 4 C (or aliquot and store at -20 C), protecting from light. When handled properly, solutions are stable for many years.

RNase A Stock Solution (1mg/ml):
To prepare, add 1 mg RNase A to 1 ml distilled water. Aliquot and store at -20 C freezer.

PI Working Solution (1 ug/ml PI and 10 ug/ml RNase A in PBS):
To prepare, add 2 ul PI Stock Solution and 20 ul RNase A Stock Solution to 2 ml PBS



Incubate sections in dark for 30 minutes at room temperature or 20 minutes at 37 C.



Staining Pattern: Nuclei will be stained red.


Suggested Use

Counterstain for immunofluorescence when green (FITC) or blue (AMCA) fluorescent marker is used.


Link Reference:
http://www.ihcworld.com/_protocols/counterstain_solutions/PI.htm