Paraffin-embedded tissue (PET) blot method: application to Alzheimer disease.
Traditionally, immunohistochemistry, immunoblotting, and histoblot have been used to detect protein in tissue samples. However, each of these techniques has a number of disadvantages. The sensitivity of protein detection in immunohistochemistry is lost due to fixation or paraffin embedding methods that modify antigenic sites. The anatomical resolution and specific cellular involvement are lost in immunoblotting. Histoblot, a hybrid of these two techniques, is able to resolve these issues, but it cannot be applied to formalin-fixed tissues. A recent technique, paraffin-embedded tissue (PET) blot, retains the superior protein detection and anatomical resolution of histoblot and is applicable to formalin-fixed tissues. Unfortunately, a major obstacle to the widespread application of PET is the lack of a detailed methodological description. In this paper, we describe a PET blotting method that was formulated from our own empirical and experimental research in Alzheimer disease and a systematic review of the current literature. From this, we conclude that PET can be applied to a variety of conditions with a wide spectrum of pathology.
Moh
Calvin F
CF
Department of Pathology, Case Western Reserve University, Cleveland, OH, USA.
Siedlak
Sandra L
SL
Tabaton
Massimo
M
Perry
George
G
Castellani
Rudy J
RJ
Smith
Mark A
MA
D000544:Alzheimer Disease / Q000378:metabolism; D005557:Formaldehyde; D006624:Hippocampus / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007150:Immunohistochemistry / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods*; D016707:Tissue Fixation / Q000379:methods
D005557:Formaldehyde