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pH3 Antibody Staining Protocol for Zebrafish
Published: Jan 20, 2012 DOI: 10.21769/BioProtoc.175 Views: 27258
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Abstract
Phosphorylation of histone H3 is tightly correlated with chromosome condensation during both mitosis and meiosis. Staining for phosphor histone H3 is a good indicator for the proliferation of the interested cells. This protocol provides a method to stain p-Histone H3 in zebrafish embryos.
Materials and Reagents
- PFA
- Phosphate buffered saline (PBS)
- Acetone
- Alcohol
- PBS-Tween 20
- H2O2
- DMSO
- Maleic Acid
- p-Histone H3 antibody, 200 μg/ml (Cell Signaling Technology, catalog number: 9701 )
- Blocking regent (Roche Diagnostics, catalog number: 1096176 )
- Diaminobenzidine (DAB) (Sigma-Aldrich, catalog number: D-5905 )
- Peroxidase-conjugated Affinity Goat anti rabbit Ig (Promega Corporation, catalog number: W4011 )
- Block solution (see Recipes)
- DAB solution (see Recipes)
- 10x MAB buffer (see Recipes)
Equipment
- Shaker
- Parafilm
- Cold-room
Procedure
Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.
Category
Cell Biology > Cell staining > Reactive oxygen species
Cell Biology > Cell signaling > Phosphorylation
Cell Biology > Cell structure > Chromosome
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