Optimized 2D Gel Protocol for Zebrafish

Based on the procedures for 2D gel electrophoresis described in [14 (link)], we optimized conditions and settings specifically for the zebrafish samples. After the last washing step of the deyolking procedure, care was taken to remove the supernatant completely in order to keep the salt concentration as low as possible. Total protein precipitation, to reduce salts and other interfering substances, is not required for analytical gels but may be applied for preparative gels. We recommend as starting material 50–100 embryos for analytical gels and 500 embryos for preparative gels.

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Link V., Shevchenko A, & Heisenberg C.P. (2006). Proteomics of early zebrafish embryos. BMC Developmental Biology, 6, 1.

Publication 2006

2d gel electrophoresis Embryos Gels Protein Salt Salts Zebrafish

Corresponding Organization :

Other organizations : Max Planck Institute of Molecular Cell Biology and Genetics

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Protocol cited in 39 other protocols

Variable analysis

independent variables

Optimized conditions and settings for zebrafish samples

dependent variables

Protein separation and visualization on 2D gel electrophoresis

control variables

Removal of supernatant after the last washing step of the deyolking procedure to keep the salt concentration as low as possible
Total protein precipitation to reduce salts and other interfering substances (may be applied for preparative gels)

controls

No positive or negative controls were explicitly mentioned.

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