Immunohistochemistry Protocol for Neural Markers

Immunostainings for glial fibrillary acidic protein (ZRF1 from DSHB, 1:200), choline O-acetyltransferase (AB144P from Millipore, 1:500) and protein kinase C alpha (SC-208 from Santa-Cruz, 1:500) were performed after completion of the FISH protocol according to de Oliveira-Carlos et al., 2013 (link). For chat, antigens were retrieved in preheated 10 mM sodium citrate buffer for 6’ at 85 °C. Sections were washed in PBS and 0.3% PBSTx prior to primary antibody incubation. Following the protocol of de Oliveira-Carlos et al., 2013 (link), we washed the sections three times in PBSTx, and incubated in anti-goat or anti-rabbit IgG (H+L) Alexa 488-conjugated secondary antibodies (Invitrogen, 1:750).

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Indrischek H., Hammer J., Machate A., Hecker N., Kirilenko B., Roscito J., Hans S., Norden C., Brand M, & Hiller M. (2022). Vision-related convergent gene losses reveal SERPINE3’s unknown role in the eye. eLife, 11, e77999.

Publication 2022

AB144P SC-208

Anti igg Antibodies Antibody Antigens Buffer Choline o acetyltransferase De protocol Fish Glial fibrillary acidic protein Goat Protein kinase c alpha Rabbit Sodium citrate

Corresponding Organization :

Other organizations : Senckenberg Research Institute and Natural History Museum Frankfurt/M, Max Planck Institute of Molecular Cell Biology and Genetics, Max Planck Institute for the Physics of Complex Systems, Center for Systems Biology Dresden, LOEWE Centre for Translational Biodiversity Genomics, Goethe University Frankfurt, TU Dresden, VIB-KU Leuven Center for Brain & Disease Research, KU Leuven, Supply Chain Competence Center (Germany)

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Variable analysis

independent variables

Immunostainings for glial fibrillary acidic protein (ZRF1 from DSHB, 1:200)
Immunostainings for choline O-acetyltransferase (AB144P from Millipore, 1:500)
Immunostainings for protein kinase C alpha (SC-208 from Santa-Cruz, 1:500)

dependent variables

Not explicitly mentioned

control variables

Sections were washed in PBS and 0.3% PBSTx prior to primary antibody incubation
Sections were washed three times in PBSTx, and incubated in anti-goat or anti-rabbit IgG (H+L) Alexa 488-conjugated secondary antibodies (Invitrogen, 1:750)

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

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