Immunohistochemistry of Zebrafish Hair Cells

Fish used for immunohistochemistry were fixed for either 2 h at room temperature or overnight at 4°C in 4% paraformaldehyde. Antibody labeling was carried out as previously described (Stawicki et al., 2014 (link)). Fish used for hair cell counts were labeled with either a mouse anti-parvalbumin primary antibody (Millipore, MAB1572) diluted at 1:500 in antibody block (5% heat-inactivated goat serum in PBS, 0.2% Triton, 1% DMSO and 0.2% BSA) or a rabbit anti-parvalbumin primary antibody (Thermo-Fisher Scientific, PA1-933) diluted at 1:1000 in antibody block.

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Stawicki T.M., Linbo T., Hernandez L., Parkinson L., Bellefeuille D., Rubel E.W, & Raible D.W. (2018). The role of retrograde intraflagellar transport genes in aminoglycoside-induced hair cell death. Biology Open, 8(1), bio038745.

Publication 2018

MAB1572

5 heat Anti antibody Antibody Antibody block Dmso Fish Goat Hair Immunohistochemistry Mouse Paraformaldehyde Parvalbumin Rabbit Serum

Corresponding Organization :

Other organizations : Lafayette College, University of Washington

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Variable analysis

independent variables

Duration of fixation: 2 h at room temperature or overnight at 4°C

dependent variables

Antibody labeling
Hair cell counts

control variables

4% paraformaldehyde used for fixation
Antibody labeling protocol previously described (Stawicki et al., 2014)
Mouse anti-parvalbumin primary antibody diluted 1:500 in antibody block
Rabbit anti-parvalbumin primary antibody diluted 1:1000 in antibody block

controls

No positive or negative controls explicitly mentioned

Annotations

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