Double immunohistochemistry with horseradish peroxidase and alkaline phosphatase detection systems.
We describe here a protocol optimized for formalin-fixed paraffin-embedded tissue sections that enables the detection of two antigens. This technique allows immunohistochemistry to be performed with detection systems allowing observation by light microscopy. This chapter discusses the choice of appropriate protocols as well as the choice of visualization systems.In doing so, we provide examples of representative results obtained with this protocol and describe necessary controls; additionally, we discuss common problems associated with this methodology, and detail troubleshooting recommendations. Although this method has been optimized for liver sections, it may be applicable for performing double immunostaining in a variety of tissue samples.
Sarrazy
Vincent
V
Faculté deMédecine et de Pharmacie, EA 3842, Institut Fédératif de Recherche 145, Université de Limoges, Limoges, France.
Desmoulière
Alexis
A
D000469:Alkaline Phosphatase / Q000032:analysis*; D000818:Animals; D006735:Horseradish Peroxidase / Q000032:analysis*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D066298:In Vitro Techniques
D006735:Horseradish Peroxidase; D000469:Alkaline Phosphatase