Antigen retrieval heating conditions

The efficacy of the antigen retrieval procedure also depends on the treatment duration and the cooling conditions (see Subheadings 3.1.-3.5.), The power of the heating device and the buffer volume are among the major factors that determine the duration of a particular HIER protocol. 

The staining protocol and differences in pretreatment methods are an important factor when formalin-fixed and paraffin-embedded material is nsed, as not all antibodies work satisfactorily under these conditions. Several different approaches can be found in the literature. Heat-induced antigen retrieval by the use of microwave heat treatment has been demonstrated to be more effective in labeling intraneuronal Ap compared to no or enzymatic pretreatment [29]. Information on the use of FA is inconsistent. It is widely used to increase the immunoreactiv-ity of amyloid plaques in AD however, it has been shown that the effect on intraneuronal Ap is low and that use of FA sometimes results in a loss of cellular detail [30]. A recent study tried to quantify the effect of FA pretreatment and analyzed plaque load and intraneuronal Ap immunoreactivity using an APi, antibody in 6-month-old APP/PSIKI mice (Fig. 3 reproduced from [25] with permission from Elsevier). 

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