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Fluorescent immunolabeling of embryonic kidney samples
1Department of Genetics and Development, Columbia University, New York, NY, USA. cc2636@columbia.edu
Methods in Molecular Biology (Clifton, N.J.)
|May 29, 2012
Summary
This chapter details a simple fluorescent immunolabeling protocol for embryonic kidney samples. It enables detection of key kidney development proteins and serves as a foundation for new antibody optimization.
Area of Science:
- Developmental Biology
- Cell Biology
- Immunohistochemistry
Background:
- Fluorescent immunolabeling is crucial for visualizing cellular structures and protein localization in biological samples.
- Embryonic kidney development involves complex cellular processes and requires precise molecular detection methods.
Purpose of the Study:
- To provide a basic, adaptable fluorescent immunolabeling protocol for embryonic kidney tissues.
- To establish a reliable method for detecting established kidney development markers.
Main Methods:
- The protocol involves five key steps: permeabilization, primary antibody incubation, washes, secondary antibody incubation, and final washes.
- Adaptable for various sample types including thin sections and whole mounts by adjusting incubation parameters and buffer composition.
Main Results:
- Successfully and consistently detected key kidney development proteins such as CalbindinD-28k, E-cadherin, Pax2, podocalyxin, α-SMA, and Phospho-Histone H3.
- Demonstrated protocol efficacy across different sample preparations.
Conclusions:
- This protocol offers a straightforward and versatile method for fluorescent immunolabeling in embryonic kidney research.
- It serves as an excellent starting point for optimizing immunolabeling of novel antibodies in kidney tissues.
