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Sections were either stained with hematoxylin and eosin (H&E) or boiled for 1 minute for antigen retrieval.
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The slides were heated in sodium acetate solution at 95°C for 10 minutes for antigen retrieval.
Then the sections were incubated with 2N HCl solution for 20 minutes, for antigen retrieval.
Prior to staining, slices were incubated in 99% formic acid for 7 minutes for antigen retrieval; this procedure is known to enhance antibody recognition of Amyloid beta [16].
The slides were placed in a pressure cooker with citrate buffer for 30 minutes for antigen retrieval, prior to rinsing with de-ionized water.
The sections were pretreated with 0.1 M citrate buffer in a pressure cooker for 2.5 minutes for antigen retrieval.
The sections were then boiled under pressure in citrate buffer (pH 6.0) for 5 minutes for antigen retrieval.
Sections were boiled in 10 mM sodium citrate buffer (pH 6.0) for 10 minutes for antigen retrieval.
Following deparaffinisation, sections were incubated in a solution of 10 μg/mL proteinase K (Qiagen, Limberg, the Netherlands) at room temperature for 20 minutes for antigen retrieval.
Afterward the slides were deparaffinized, hydrated, placed in 10 mM citrate buffer (pH 6) and microwaved for a total of 20 minutes for antigen retrieval.
Additional sections were manually deparaffinized in xylene, rehydrated in a series of graded ethanol solutions, boiled in10 mM sodium citrate buffer (pH 6.0) for ten minutes, then cooled for 20 minutes for antigen retrieval.
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