PUBLICATION
Protein Purification and Western Blot Detection from Single Zebrafish Embryo
- Authors
- Schnabel, D., Castillo-Robles, J., Lomeli, H.
- ID
- ZDB-PUB-190814-14
- Date
- 2019
- Source
- Zebrafish 16(6): 505-507 (Journal)
- Registered Authors
- Lomeli, Hilda, Schnabel, Denhi
- Keywords
- protein purification, western blot, zebrafish embryo
- MeSH Terms
-
- Animals
- Blotting, Western/methods*
- Embryo, Nonmammalian/chemistry*
- Fish Proteins/isolation & purification*
- Genotyping Techniques/methods*
- Zebrafish*/genetics
- Zebrafish*/growth & development
- PubMed
- 31408407 Full text @ Zebrafish
Citation
Schnabel, D., Castillo-Robles, J., Lomeli, H. (2019) Protein Purification and Western Blot Detection from Single Zebrafish Embryo. Zebrafish. 16(6):505-507.
Abstract
Characterization of a protein of interest during development is essential for functional studies. A general strategy for understanding the function of a particular protein involves the generation of null mutations, or treatment with drugs, that interfere with its activity. To demonstrate that the synthesis, stability, or activity of a protein has been affected, accurate and efficient detection of low amounts of protein is essential. This can be achieved by immunohistochemistry or by western blot. Here we describe a method for the detection of proteins from single de-yolked zebrafish embryos. This procedure includes a fixation step and the concomitant elimination of lipids from the yolk cell. We show that this approach allows the rapid analysis of proteins in embryos without having to manually remove the yolk. This method provides a convenient alternative for genotyping of mutant embryos as early as the 128 cell stage. In addition, in drug- or morpholino-treated embryos, the correlation between the penetrance of a phenotype and the concentration of a protein present may be established.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping