Membranipora in situ + mapk, a re-try

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Since I saved some de-glycerolled embryos I’ll repeat the immuno staining with a longer developing time.

gata b nk2.1
  • Washed 1h in PBT (4x).
  • Washed 3h in 5% NGS (3x).
  • Added anti-mapk antibody 1:100.
  • Let overnight at 4°C.


  • Washed primary antibody with 3x PBT 5min + 3x 30min.
  • 5% NGS 3x 30 min.
  • Added both anti-mouse-pod/hrp at 1:250 for 4h at room temp.
  • Washed 4x 5min with PBT and let overnight at 4°C.


  • Wash 1x PBT.
  • Wash 3x TNT buffer.
  • Develop with cy3 for 5min.
  • Wash with PBS (no detergent solution).
  • Embed in TDE.