Stain-Free Approach for Western Blot Analysis of Zebrafish Embryos

Western blots are employed to detect and characterize amounts of proteins in biological samples. Quantifications are traditionally carried out through data normalization by housekeeping protein method. This approach does not account for variations not intrinsically dependent on the sample such as different experimental conditions and type of samples. Zebrafish researchers often face the challenge of comparing embryos at different developmental stages or from different strains. Housekeeping protein amount can change in these conditions therefore adding an unwanted quantification error. Here we describe the method to analyze mutant zebrafish embryos at different stages by western blot using the Stain-Free technology for normalization. We present Globin X quantification at 2 and 5 days postfertilization in wild type and in the bloodless Vlad Tepes (vlt) zebrafish mutant that lack red blood cells.