I encountered some problems during the western blotting of protein extracted from zebrafish larvae. I use the chloroform: methanol=1:3 configuration to remove the yolk solution, but after centrifugation, an insoluble precipitate is formed, resulting in poor protein quality. I would like to ask how to remove the yolk efficiently without affecting the quality of the protein. thank you!
We had success using the method in here:
Article Targeted deletion of the zebrafish actin-bundling protein L-...
Deyolking embryo homogenates for running on gels is a perennial problem. We've used everything from the old-school titurating with a p200
Article Proteomics of early zebrafish embryos
to the heptane/MeOH method published by Schnable Article Protein Purification and Western Blot Detection from Single ...
Especially for later stages, the best method is to mechanically deyolk by drawing the embryos through a glass transfer pipette that has been drawn out in a flame and broken open such that the opening is narrower than the yolk ball but wider than the embryo's head.
If you come up with a better approach, you should publish it.
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