Immunodetection of 4-HNE adducts was conducted as previously described (Roychowdhury et al., 2009b (link)). Liver tissue was formalin-fixed and paraffin-embedded, after which it was sectioned and mounted. Immunohistochemistry was then performed using rabbit antisera against 4-HNE (Alpha diagnostic 1:100). Following primary incubation, sections were conjugated using Vectastain Elite Rabbit IgG Kit (Vector Labs) and visualized using DAB substrate chromogen. Following development, slides were then counterstained using Gill’s hematoxylin (Vector labs). Images were acquired on a 20X objective by a blinded investigator.
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Immunodetection of 4-HNE Adducts in Liver
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Antisera
Chromogen
Diagnostic
Formalin
Gill
Hematoxylin
Immunohistochemistry
Liver
Paraffin
Rabbit
Tissue
Vector
Corresponding Organization : Alcohol Research Group
Other organizations : Cleveland Clinic Lerner College of Medicine
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Variable analysis
independent variables
- Immunodetection of 4-HNE adducts
- Visualization of 4-HNE adducts in liver tissue
- Formalin-fixed and paraffin-embedded liver tissue
- Sectioning and mounting of liver tissue
- Primary incubation with rabbit antisera against 4-HNE (Alpha diagnostic 1:100)
- Conjugation using Vectastain Elite Rabbit IgG Kit (Vector Labs)
- Visualization using DAB substrate chromogen
- Counterstaining with Gill's hematoxylin (Vector labs)
- Acquisition of images on a 20X objective by a blinded investigator
- Not explicitly mentioned
- Not explicitly mentioned
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