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: Muscle, Myotendinous junction, Antibody
Materials and Reagents
Equipment
Procedure
permeabiliza-tion®
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
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We buy phalloidin from Life Technologies and dissolve it in 1.5 mL MeOH. We then aliquot and freeze the phalloidin stock at -20 C. Before incubating the zebrafish embryos in phalloidin, it is diluted 1 part phalloidin to 19 parts PBS with 2% Triton-X 100.
PBS with 2 % trinton X100 menas? can you elaborate
There is a recipe for PBS with 2% TritonX 100 in the protocol above.