Immunofluorescent staining on paraffin-embedded tissue sections

 
LEAD AUTHOR

Marcus Parrish, Doctoral Candidate, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA USA

mparrish@mit.edu

OTHER AUTHORS:

Guanyu Gong, Postdoctoral Researcher, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

Jenny Kay, Doctoral Candidate, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA

Bevin Engelward, Professor, Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA


PROTOCOL TYPE:
Protocol Document
SYNOPSIS:

This protocol describes the steps necessary to fluorescently detect proteins in paraffin-embedded tissue sections. After depariffinizing and rehydrating the tissues, antigens are exposed through incubation with boiling antigen retrieval buffer. Subsequently, sections are blocked with bovine serum albumin and probed with primary antibody overnight. After a series of washes, the sections are incubated with secondary antibody and detected with a fluorescent microscope.


 
PUBLICATIONS:

Rai P, Parrish M, Tay IJ, Li N, Ackerman S, He F, Kwang J, Chow VTK, Engelward BP, Streptococcus pneumoniae secretes hydrogen peroxide leading to DNA damage and apoptosis in lung cells, Proc Natl Acad Sci U S A. 2015; 112:E3421-30. doi: 10.1073/pnas.1424144112. PubMed PMID: 26080406.

Li N, Parrish M, Chan TK, Yin L, Rai P, Yoshiyuki Y, et al. Influenza infection induces host DNA damage and dynamic DNA damage responses during tissue regeneration. Cell Mol Life Sci. 2015;72(15):2973-88. doi: 10.1007/s00018-015-1879-1. PubMed PMID: 25809161.


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