Transfer and multiplex immunoblotting of a paraffin embedded tissue.

Abstract:

:As we transition from genomics to the challenges of the functional proteome, new tools to explore the expression of proteins within tissue are essential. We have developed a method of transferring proteins from a formalin fixed, paraffin embedded tissues section to a stack of membranes which is then probed with antibodies for detection of individual epitopes. This method converts a traditional tissue section into a multiplex platform for expression profiling. A single tissue section can be transferred to up to ten membranes, each of which is probed with different antibodies, and detected with fluorescent secondary antibodies, and quantified by a microarray scanner. Total protein can be determined on each membrane, hence each antibody has its own normalization. This method works with phospho-specific antibodies as well as antibodies that do not readily work well with paraffin embedded tissue. This novel technique enables archival paraffin embedded tissue to be molecularly profiled in a rapid and quantifiable manner, and reduces the tissue microarray to a form of protein array. This method is a new tool for exploration of the vast archive of formalin fixed, paraffin embedded tissue, as well as a tool for translational medicine.

journal_name

Proteomics

journal_title

Proteomics

authors

Chung JY,Braunschweig T,Baibakov G,Galperin M,Ramesh A,Skacel M,Gannot G,Knezevic V,Hewitt SM

doi

10.1002/pmic.200401343

subject

Has Abstract

pub_date

2006-02-01 00:00:00

pages

767-74

issue

3

eissn

1615-9853

issn

1615-9861

journal_volume

6

pub_type

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