Vectastain anti rabbit hrp kit

Paraffin sections were deparaffinized and antigens were unmasked with citrate (pH 6) and heat. Sections were treated with 3% hydrogen peroxide before being blocked with 1% goat serum in PBS. Samples that were unmasked with citrate buffer were stained with phospho-Histone H3, and cleaved caspase 3. Sections were then stained using a Vectastain anti-rabbit HRP kit (Vector Laboratories) and a DAB (3,3’-diaminobenzidine) peroxidase substrate kit (Vector Laboratories). Briefly, sections were then incubated with biotinylated anti-rabbit antibodies, then with streptavidin-HRP and developed with DAB. Slides were scanned at the NYULMC Experimental Pathology Research Laboratory (NYULMC, New York, NY, USA) and whole tissue staining was quantified by Aperio ImageScope software (Leica Biosystems) in non-necrotic areas.

Paraffin sections were deparaffinized and antigens were unmasked with citrate (pH 6) and heat. Sections were treated with 3% hydrogen peroxide in Methanol before being blocked with 1% goat serum in PBS. Samples that were unmasked with citrate buffer were stained with phospho-H3, cleaved caspase 3, PGP9.5, Th, Trpv1, VAchT,pan-Trk, and phospho-eIF2α. Sections were then stained using Vectastain anti-rabbit HRP kit (Vector Laboratories) and DAB (3,3’-diaminobenzidine) peroxidase substrate kit (Vector Laboratories). Briefly, sections were then incubated with biotinylated anti-rabbit antibodies, then by streptavidin-HRP and developed with DAB. Slides were scanned at the NYULMC Experimental Pathology Research Laboratory (NYULMC, New York, NY, USA) and whole tissue staining was quantified by Aperio ImageScope software (Leica Biosystems) in non-necrotic areas.
Human PDAC TMAs (IRB protocol: LAB05–0854) were stained for PHGDH at the NYULMC Center for Biospecimen Research and Development laboratory. TMAs also were stained with PGP9.5 as described above. Tissues were scored from 0 to 3 for PHGDH intensity by a GI pathologist or quantified by Aperio ImageScope software.