Fluorescent immunohistochemistry and in situ hybridization analysis of mouse pancreas using low-power antigen-retrieval technique.
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 is always required for different tissues and antigens. In this study, by using a low-power antigen-retrieval technique with appropriate dilution of antibodies, we successfully immunostained key antigens in pancreas such as insulin, PDX-1, glucagon, cytokeratin, and CD31, which have previously presented a particular challenge for investigators because of the rapid autodigestion and high nonspecific antibody binding in this tissue. Satisfactory results were obtained when immunohistochemistry and fluorescence in situ hybridization analysis were combined in the same slides.
Ge
Shundi
S
Division of Research Immunology/Bone Marrow Transplantation, Childrens Hospital Los Angeles, Los Angeles, CA 90027, USA.
Crooks
Gay M
GM
McNamara
George
G
Wang
Xiuli
X
D000818:Animals; D005455:Fluorescent Antibody Technique; D005934:Glucagon / Q000378:metabolism*; D018398:Homeodomain Proteins / Q000378:metabolism*; D017404:In Situ Hybridization, Fluorescence; D007328:Insulin / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism*; D051379:Mice; D016513:Mice, SCID; D010179:Pancreas / Q000378:metabolism*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000378:metabolism*; D015534:Trans-Activators / Q000378:metabolism*
D018398:Homeodomain Proteins; D007328:Insulin; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D015534:Trans-Activators; C084010:pancreatic and duodenal homeobox 1 protein; D007633:Keratins; D005934:Glucagon