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J Immunol Methods. 2010 Jul 31;359(1-2):56-60. doi: 10.1016/j.jim.2010.05.002. Epub 2010 May 12.

A method for extracting tissue proteins for use in lymphocyte function assays.

Author information

1
Immunology and Diabetes Unit, St. Vincent's Institute of Medical Research, 9 Princes St, Fitzroy, Victoria 3065, Australia.

Abstract

Currently human T-cell responses to unpurified tissue extracts cannot be easily measured because current cell lysis methods yield a lysate that is toxic. Here we describe the optimization of a protocol for extracting proteins from human tissues in a format that is compatible with the functional analysis of human T cells. The tissue was homogenized in a mixture of butan-1-ol, acetonitrile and water and then lyophilized. Lyophilized protein extracts were dissolved in 8M urea because urea did not affect T-cell function when present at 0.08 M or less. Using this method cytokine production and proliferation responses were detected from islet, acinar and spleen extracts. Hence, our method allows the rapid preparation of human tissue lysates in a format that is compatible with the analysis T-cell responses. We suggest that this method will facilitate the analysis of adaptive immune responses to tissues in transplantation, cancer and autoimmunity.

PMID:
20470778
DOI:
10.1016/j.jim.2010.05.002
[Indexed for MEDLINE]

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