About this item:

143 Views | 87 Downloads

Author Notes:

E-mail Address : dfalls@emory.edu.

We thank Julie Siegenthaler and Dr. Michael Miller (State University of New York, Syracuse, NY) for discussions.

Subjects:

Research Funding:

This work was supported by National Institute on Deafness and Other Communication Disorders Grant RO1 DC03190 (M.B.L.).

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Neurosciences
  • Neurosciences & Neurology
  • CELLS
  • PROLIFERATION
  • MIGRATION
  • IMMUNOHISTOCHEMISTRY
  • FOREBRAIN

Antigen-retrieval procedure for bromodeoxyuridine immunolabeling with concurrent labeling of nuclear DNA and antigens damaged by HCl pretreatment

Tools:

Journal Title:

Journal of Neuroscience Nursing

Volume:

Volume 27, Number 22

Publisher:

, Pages 5837-5844

Type of Work:

Article | Final Publisher PDF

Abstract:

Visualizing the proliferation marker bromodeoxyuridine (BrdU) requires pretreatment of tissue, typically with dilute hydrochloric acid (HCl). We report here that pretreatment by steam heating of paraformaldehyde-fixed tissue sections covered with citrate buffer yields much brighter labeling of BrdU than HCl pretreatment, allows labeling with many antibodies greatly superior to HCl pretreatment, and allows concurrent high-quality labeling of nuclei with the DNA-binding dyes Hoechst, DAPI (4′,6-diamidino-2-phenylindole), and Syto24. Standard use of antigen retrieval by steamer can facilitate new insights into mechanisms regulating normal progenitor and tumor cell proliferation and novel understandings of protein expression through increased sensitivity of immunohistochemical analysis.

Copyright information:

Copyright © 2007 Society for Neuroscience.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 4.0 International License (https://creativecommons.org/licenses/by/4.0/).
Export to EndNote