Fluorescent Immunohistochemistry and In Situ Hybridization Analysis of Mouse Pancreas Using Low Power Antigen Retrieval Technique

Shundi Ge ¹, Gay M. Crooks ^1*, George McNamara ¹ and Xiuli Wang ¹

¹ Division of Research Immunology/BMT, Childrens Hospital Los Angeles, Los Angeles, California (SG,GMC,XW), and Division of Cancer Immunotherapeutics and Tumor Immunology, City of Hope National Medical Center, Duarte, California (GM)

^* To whom correspondence should be addressed. E-mail: gcrooks{at}chla.usc.edu.

Submitted on December 14, 2005
Accepted on 2 March 2006

	Abstract

To facilitate the immunological reaction of antibodies with antigens in fixed tissues, it is necessary to unmask or retrieve the antigens through pretreatment of the specimens. However, adjustment of heating induced antigen retrieval (HIAR) is always required for different tissues and antigens. In this study, by using a low power AR (LAR) technique with appropriate dilution of antibodies, we successfully immuno-stained key antigens in pancreas such as insulin, PDX-1, glucagon, cytokeratin, and CD31, which have presented a particular challenge for investigators in the past, because of the rapid autodigestion and high nonspecific antibody binding in this tissue. Satisfactory results were obtained when immunohistochemtry and fluorescence in situ hybridization analysis was combined in the same slides.
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