CD39 Antibody – Guinea Pig

Recommended Dilution: 1:1000

Antiserum was used on perfusion fixed tissue. Perfusion: 1) calcium-free Tyrode’s solution, 2) paraformaldehyde-picric acid fixative, and 3) 10% sucrose in PBS as a cryo-protectant. Desired tissues were dissected and stored overnight in 10% sucrose in PBS.

Slide-mounted tissue sections were processed for indirect immunofluorescence. Slides were incubated with blocking buffer for 1 hour at room temperature. Primary antiserum was diluted with blocking buffer to the appropriate working concentration. Blocking buffer was removed and slides were incubated for 18-24 hours at 4ºC with primary antiserum. Slides were rinsed 3 times and then incubated with secondary antibodies for 1 hour at room temperature. Slides were again rinsed 3 times and coverslipped. Staining was examined using fluorescence microscopy.

Note: Sodium azide (NaN₃) interferes with peroxidase reactions and should not be used with peroxidase methodologies. If sodium azide is present in any steps of the staining procedure, the tissue should thoroughly be rinsed with sodium azide-free buffer before performing the peroxidase reaction.

Dilutions listed only as a recommendation. Optimal dilution should be determined by investigator.