Antibody Staining of Scale Melanocytes: A Technique to Detect Transcription Factors Expressed in Dorsal Scale-associated Melanocytes of Zebrafish Models
This video describes the antibody staining method to detect transcription factors expression in dorsal scale-associated melanocytes of zebrafish models. This immunostaining technique helps specifically identify the melanocyte-inducing transfection factor-A in the transgenic zebrafish.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. Antibody Staining of Scale Melanocytes Make up PO4 buffer with 80 mL 0.1M Na2HPO4 and 20 mL 0.1M NaH2PO4. Make sure the pH is 7.3. Make up 2x fix buffer containing 8.0g sucrose, 0.15 mL 0.2M CaCl2, and 90 mL 0.2M PO4 buffer, pH 7.3. I…
Representative Results
Figure 1. Antibody staining of scale melanocytes. A) Scales being plucked from an anesthetized miniCoopR-EGFP zebrafish. B) Unbleached scale with pigmented melanocytes and C) bleached scale from miniCoopR-EGFP zebrafish. Scale bar = 100 μM. Unbleached scale stained with a D) Mitfa antibody and <st…
Antibody Staining of Scale Melanocytes: A Technique to Detect Transcription Factors Expressed in Dorsal Scale-associated Melanocytes of Zebrafish Models. J. Vis. Exp. (Pending Publication), e20364, doi: (2023).