Michel’s fixative for immunofluorescence

Contributed by Martin Fitzpatrick, University of Birmingham, United Kingdom

Michel’s fixative for immunofluorescence

• Pour 100mls distilled water into a suitable container
• Add 0.81g potassium citrate
• Add 0.0625g N-ethylmaleimide (Handle with care)
• Add 0.123g magnesium sulphate to prepare stock solution.
• Before use add 55g ammonium sulphate and mix thoroughly to dissolve.
• Adjust pH to 7.0-7.2 with 1M KOH.
• Place tissue in fixative for 24-48hours
• Wash tissues in buffer 3x for 10 minutes
• Freeze tissues at -70’C until ready for use

This method is based, with permission, on an original protocol available here.