E-Book Overview
Matthew Guille assembles a hands-on collection of basic and essential molecular and embryological techniques for studying Xenopus and zebrafish. Easily reproducible and designed to succeed, these detailed methods include cellular techniques, techniques for the quantitative and spatial analysis of mRNA and proteins, and techniques for the expression of gene products in embryos. More specialized methods enable users to analyze promoters and transcription factors during early development, and include gel shift assays, as well as in vitro and in vivo footprinting. Molecular Methods in Developmental Biology: Xenopus and Zebrafish affords newcomers rapid access to a wide variety of essential, state-of-the-art techniques in developmental studies, and offers experienced researchers time-saving tips and new techniques from experts who have fine-tuned them for best results.
E-Book Content
Methods in Molecular Biology
TM
VOLUME 127
Molecular Methods in Developmental Biology Xenopus and Zebrafish Edited by
Matthew Guille
HUMANA PRESS
Animal Cap Assay
1
1 The Animal Cap Assay Jeremy Green 1. Introduction Over the last 10 years, the animal cap of the Xenopus laevis embryo has proved to be a versatile test tissue for a variety of molecules involved not only in animal development but also vertebrate cell regulation in general. These molecules include growth factors (1–3), cell surface receptors (4–6), signal transduction molecules (7,8), transcription factors (9), and extracellular matrix molecules (10). The “animal cap assay” provides a simple, quick, inexpensive, and quantitative bioassay for biological activity of both cloned genes and purified or unpurified proteins. The animal cap is a region of the Xenopus blastula and early gastrula stage embryo (6–12 h after fertilization). It is “animal” because the upper, pig