Published: Vol 3, Iss 11, Jun 5, 2013 DOI: 10.21769/BioProtoc.786 Views: 27969
Reviewed by: Anonymous reviewer(s)
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Abstract
Fluorescent conjugated Phalloidin is a stain that allows for visualization of F-actin. In immunohistochemistry, primary antibodies and fluorescent conjugated secondary antibodies can be used to visualize subcellular localization and relative amounts of proteins of interest. Here is a protocol for Phalloidin and antibody staining of zebrafish embryos 5 days old and younger.
Keywords: MuscleMaterials and Reagents
Equipment
Procedure
Name | Company | Product # | Dilution | Mono/polyclonal | Works withphalloidin® | Extra permeabiliza-tion® |
Beta-Dystroglycan | Novocastra | NCLbDG | 1:50 | Monoclonal | Yes | No |
F59 | Developmental StudiesHybridoma Bank (DSHB) | 1:10 | Monoclonal | Yes | No | |
Fibronectin | Sigma | F3648 | 1:50 | Polyclonal | Yes | No |
Laminin-111 | Thermo Scientific | RB-082-A0 | 1:50 | Polyclonal | Yes | No |
Paxillin | BD TransductionLaboratories | 610051 | 1:50 | Monoclonal | Yes | No |
FAK pY397 or pY861 | Invitrogen | 1:50 | Polyclonal | Yes | No | |
Dystrophin | Sigma | D8043 | 1:50 | Monoclonal | Yes | No |
MF20 | DSHB | 1:10 | Monoclonal | Not well | No | |
F310 | DSHB | 1:10 | Monoclonal | Not well | No | |
Vinculin | Sigma | V4505 | 1:10 | Monoclonal | Unknown | PBS 2% TritonX-100® for 2.5 hours at room temp |
4D9/engrailed | DSHB | 1:2 | Monoclonal | Yes | 10 minutes in acetoneat -20 °C | |
Beta-catenin | Abcam | Ab6302 | 1:500 | Monoclonal | Unknown | Requires specialfixative (4% PFA, 4% sucrose, 3 mM CaCl2, 1x PBS) |
GFP | Molecular Probes | A21311 | 1:200 | Polyclonal | Unknown | 2 h fix in 4% PFAat room temp |
Recipes
Acknowledgments
This protocol was adapted from the previous publications: Goody et al. (2010) and Goody et al. (2012). Development of this protocol was supported by NIH grant RO1 HD052934-01A1 to CAH. MFG would like to thank the University of Maine Graduate School of Biomedical Sciences and Engineering for funding.
References
Article Information
Copyright
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Goody, M. F. and Henry, C. A. (2013). Phalloidin Staining and Immunohistochemistry of Zebrafish Embryos. Bio-protocol 3(11): e786. DOI: 10.21769/BioProtoc.786.
Category
Developmental Biology > Morphogenesis
Cell Biology > Cell imaging > Fluorescence
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