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8271868 
Journal Article 
Immunocytochemistry as a tool for zebrafish developmental neurobiology 
Novak, AE; Ribera, AB; , 
2003 
1 
Methods in Cell Science
ISSN: 1381-5741
EISSN: 1573-0603 
79-83 
English 
14739591 
Two methods are presented here that allow clear visualization of antibody localization in zebrafish whole mount preparations, both for immunocytochemistry (ICC) alone and in combination with in situ hybridization (ISH). The first protocol describes ICC performed using a modified permeabilization technique and the chromogen AEC (3-Amino-9-ethylcarbazole). The second protocol describes the co-localization of transcriptional and translational products using a combined ISH/ICC protocol. A fluorescing chromogen (Fast Red, FR) is used to detect mRNA transcripts by ISH, and is combined with ICC that uses a secondary antibody conjugated to a different fluorescent molecule (Alexa 488). These procedures allow the identification of gene expression patterns in cell types identifiable with known antibodies.