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2 protocols using anti p16 sc 56330

1

HPV Status Evaluation by p16 Immunostaining

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HPV status was evaluated by p16 immunostaining of paraffin-embedded tissue samples. Anti-p16 (sc-56330; Santa Cruz Biotechnology Inc., Santa Cruz, CA, USA) was used as the primary antibody and the VECTASTAIN Elite ABC kit (PK-6100; Vector Laboratories Inc., Burlingame, CA, USA) was used as the secondary antibody. ImmPACT DAB (SK-4105; Vector Laboratories Inc.) was used as the color developing agent. The definition of p16-positive was staining of >70% of the cancer nucleus or cytoplasm, and this was judged by two double-blinded doctors.
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2

Western Blot Analysis of Neurodegeneration

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Cell lysates and SNc tissues were homogenized in RIPA lysis buffer, and protein concentration was determined by the Bradford assay (Bio-Rad, Hercules, CA, USA). A 30-μg protein aliquot of each sample was separated using standard SDS-PAGE and transferred onto a PVDF membrane (Millipore, Bedford, MA). Immunoreactive bands were detected by enhanced chemiluminescence plus detection reagent (Pierce, Rockford, IL) and analyzed using the ImageQuant™ LAS 4000 imaging system (GE Healthcare, Pittsburgh, PA, USA). The following primary antibodies were used: anti-TH (T1299, Sigma, St Louis, MO, USA), anti-p16 (sc-56330, Santa Cruz Biotechnology, USA), anti-β-actin (BM0627, Boster, Pleasanton, CA, USA), anti-LC3 (3868, Cell Signaling Technology, USA), anti-p62 (23214, Cell Signaling Technology, USA), anti-COX IV (4844, Cell Signaling Technology, USA), anti-TOM20 (42406, Cell Signaling Technology, USA), anti-Parkin (ab77924, Abcam, USA), and anti-PINK1 (ab23707, Abcam, USA).
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