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.
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.