Paraffined blocks of hepatic tissue were sectioned and deparaffinized with xylene and subsequently rehydrated with alleviated concentrations of alcohol. The sections were left for 10 min in 3% H2O2 followed by 30 min in 0.1% trypsin at 37 °C, for antigen retrieval. They were subsequently incubated overnight at 4 °C with antibodies of polyclonal rabbit against Bcl2, NF-κB (p65), Bax, PPAR-γ, and caspase-3 (1:1000 dilution; Abcam, Cambridge, UK). Sections were treated at 37 °C for 30 min with biotinylated goat anti-rabbit (Invitrogen), the secondary antibody, and subsequently stained with 3, 3-diaminobenzidine and eventually counterstained with hematoxylin. Afterward, image analysis was carried out via ImageJ software (version 1.48) to assess the area percent (A%).
Biotinylated goat anti rabbit
Manufactured by Thermo Fisher Scientific
Sourced in France
Biotinylated goat anti-rabbit is a secondary antibody conjugated with biotin. It is designed for use in various immunoassays and detection techniques where the identification or quantification of rabbit primary antibodies is required.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using biotinylated goat anti rabbit
1
Histological Assessment of Hepatic Biomarkers
2
Quantitative Western Blot Analysis
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Western blot was carried out as previously described49 (link). The primary antibodies used were mouse monoclonal anti-glutamine synthetase (Abcam, Cambridge, UK), anti-β-actin antibody (Sigma-Aldrich, Saint-Louis, Missouri, USA), anti-α-tubulin antibody (Santa Cruz, Dallas, Texas USA.), rabbit polyclonal anti-asparagine synthetase (Abcam), anti-4E-BP1, anti-4E-BP1-pT70, anti-Erk1/2, anti-Erk1/2-pT202/Y204, anti-elF-2α-pS51, anti-elF-2α (Ozyme, Saint Quentin Yvelines, France), and anti-BCL2A1 antibody (Abcam). Secondary antibodies were as follows: biotinylated rabbit-anti mouse (Dako, Courtaboeuf, France) and biotinylated goat anti-rabbit (Invitrogen, Carlsbad, CA, USA). Detection was performed using Luminata Forte Western HRP substrate (Millipore Corporation, Billerica, MA). Quantifications were carried out by densitometric analysis using Kodak software as previously described49 (link). β-actin or α-tubulin was used as loading control.
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