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Purified anti-human CD279 (PD-1) Antibody
329902
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I wanted to back up some flow cytometry data from a colleague by studying TFH cells with immunofluorescence. I use this antibody at a 1 in 200 dilution in PBS (1 hour, RT) for staining my acetone fixed (20mins, 4oC, 6micron) sections. Amplification with anti IgG1 Alexa flour 647 or Alexa flour 546 works fine for me (not tried it with Alexa flour 488).
Immunofluorescence
Tonsil
1 in 200, 1 hour at RT.
Blocked tonsil with 10% normal goat serum (all secondaries were raised in goat)
45 minutes with 1/300 Goat anti mouse IgG1 alexa flour 546
Incubated with Hoescht
Zeiss Zen 780 confocal microscope.
Clear staining within the germinal centre of PD1+ CD3+ cells.
Image shows PD1 cells in red, CD20 (B cells) in green and CD3 in blue.
It works!
The staining isn't that bright so make sure you use appropriate isotype controls.
Stains PD1 in acetone fixed human tissue.