Tissue samples were fixed in 10% Formalin and embedded in paraffin wax (Leica RM2255 microtome). Sections were cut in Palm slides and underwent Haematoxylin&Eosin (H&E) staining and automated Immunohistochemistry staining (Leica ST5010). Histology slides were imaged on a Pannoramic 250 Slide Scanner (3DHISTECH) and images exported to ImageJ. Images were converted to RGB stacks and a threshold was set across all images to identify positive staining. H&E and PAX8 staining was used to identify HGSC tissue. Regions of interest (ROIs) were then manually drawn around discrete areas of HGSC tissue. The data were reported as the total area percentage of the ROI staining positive. PAX8: Abcam ab13611, 1:100; F4/80: Biorad MCA497GA, 1:1000; Adenovirus: Abcam ab8251, 1:1000; DX5: Abcam ab133557, 1:250.
Ab8251
Manufactured by Abcam
Sourced in United Kingdom
Ab8251 is a mouse monoclonal antibody that recognizes Granzyme B. Granzyme B is a serine protease that is involved in apoptosis. The antibody can be used in various applications such as Western blotting, immunohistochemistry, and ELISA.
Automatically generated - may contain errors
3 protocols using ab8251
1
Histological Analysis of HGSC Tissue
2
Production of Recombinant Adenoviral Vectors
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The linearized pAdEasy or pAdEasy-HMGA-6 DNA was transfected into the AD293 cell line, a derivative of the HEK cell line, by using ViraPack Transfection Kit (Agilent Technologies) according to the manufacturer protocol. Viral production was confirmed by immunocytofluorescence assays. Cultures of AD293 cells transfected with linearized DNA encoding native AdEasy or encoding engineered AdEasy-HMGA-6 were probed for virus production using mouse anti-hexon primary antibodies (Abcam #ab8251) and Alexa fluor-coupled donkey anti-mouse immunoglobulin G (IgG) secondary antibodies containing GFP (Invitrogen #A-21202). After 7–10 days of transfection, when cytopathic effects were visible, the primary viral stock was prepared by freezing-thawing the infected cells four times in a dry ice-methanol bath and a 37°C water bath.
3
Hexon Staining Immunohistochemistry Protocol
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