Immunofluorescence staining for an oxidative damage marker on ocular sections was performed using 4-HNE antibody specific for lipid peroxidation. Six μm sections were incubated with the anti-4-HNE antibody (Alpha Diagnostic International Inc., San Antonio, TX, USA) at 4 °C for overnight followed by a rabbit anti-rabbit IgG fluorescence-conjugated secondary antibody ( Invitrogen Corp., Waltham, MA, USA) for 1.5 h at room temperature and counterstained with DAPI.
Anti 4 hne antibody
Manufactured by Alpha Diagnostic
Sourced in United States
The Anti-4-HNE antibody is a laboratory tool used to detect the presence of 4-Hydroxynonenal (4-HNE), a marker of oxidative stress. This antibody can be utilized in various analytical techniques, such as Western blotting, immunohistochemistry, and ELISA, to measure and quantify 4-HNE levels in biological samples.
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3 protocols using anti 4 hne antibody
1
Immunofluorescence Assay for Lipid Peroxidation
2
Immunofluorescence Imaging of Oxidative Stress
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Sections were incubated with anti-4-HNE antibody (Alpha Diagnostic International Inc, San Antonio, TX, USA) at 48°C for 2 hours followed by a fluorescence-conjugated donkey anti-rabbit IgG (catalog no. A21206, Invitrogen Corp. Waltham, MA, USA) for 2 hours at room temperature and counterstained with 4',6-diamidino-2-phenylindole (DAPI) (Invitrogen, Eugene, Oregon, USA). Images were examined using a BZ-X700 All-in-One inverted fluorescence microscope with structural illumination (Keyence Corp., Itasca, IL, USA) at 20× magnification spanning the entire cortical or hippocampal regions. Quantification of the staining is described in Section 2.7 .
3
Immunofluorescence Imaging of Oxidative Stress
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