p62 (human) polyclonal antibody

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Paraffin-embedded CNS tissue sections from Tg338 mice were studied by immunohistochemistry (IHC) to evaluate the brain expression and distribution of LC3-B and p62 autophagy-related proteins. After deparaffination and rehydration, sections were pre-treated by heat-induced epitope retrieval for 20 min at 96 °C in a PTLink (Dako), using Tris-EDTA buffer (pH 9.0). Endogenous peroxidase activity was then blocked and samples were incubated for 1 h at room temperature with the following primary antibodies: mouse monoclonal anti-MAP-LC3β (1:200 dilution; G-2, sc-271625; Santa Cruz Biotechnology) and rabbit polyclonal anti-p62 (1:200 dilution; PW9860; Enzo Life Sciences). Omission of the primary antibody served as a background control for nonspecific binding of the secondary antibody. Subsequently, the enzyme-conjugated polymer EnVision (Dako EnVision anti-mouse for MAP-LC3β; Dako EnVision anti-rabbit for p62) was used as visualization system, followed by an incubation with diaminobenzidine chromogen (Dako).

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