Membranipora anti-MAPK 3h, 22h, 26h, 50h, 70h immunostaining

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Started immuno staining with freshly fixed material of Membranipora for 3h, 22h, 26h, 50h, 70h post fertilization.

Anti-MAPK 1:1000 incubated at 19h30.


Did regular washes and added secondary antibody Alexa 647 1:250.


Washed in PBT and let at 4 °C in the cold room.


Stained with phallacidin and DAPI and prepared a slide in glycerol. Both seem to be working fine, but I couldn’t see the MAPK signal under the fluorescence scope. So I need to see it under the confocal.


Checked under the confocal and they were not good enough. Phallacidin was fading quickly, DAPI was weak and there was no clear signal for the MAPK. I’ll try the original concentration and DAB next time.

MAX_Mmem DAPI PHALL MAPK A647.lif - Series014 MAX_Mmem DAPI PHALL MAPK A647.lif - Series018 MAX_Mmem DAPI PHALL MAPK A647.lif - Series044