Double fluorescence labeling was performed as described previously [30 (link)]. Amongst the stored brains after India Ink angiography, 12 brains were randomly used from groups sham (n=3), SAH (n=3), low atorvastatin group (n=3), and high atorvastatin group (n=3), respectively. The intracranial internal carotid artery (ICA) was sectioned every 200 μm. Ten micrometers thick coronal sections were cut by a cryostat and incubated overnight at 4°C with the rabbit anti-Caspase 3 (1:50; Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit anti-CHOP (1:500; Sigma-Aldrich, St. Louis, MO) antibodies, followed by incubation with appropriate fluorescence dye-conjugated secondary antibodies (Jackson ImmunoResearch, West Grove, PA). The sections were visualized with a fluorescence microscope, and pictographs were recorded and analyzed with MagnaFire SP 2.1B software (Olympus, Melville, NY).
Fluorescence dye conjugated secondary antibodies
Manufactured by Jackson ImmunoResearch
Sourced in United States
Fluorescence dye-conjugated secondary antibodies are laboratory reagents used to detect and visualize target proteins in various analytical techniques, such as Western blotting, immunohistochemistry, and flow cytometry. These antibodies are designed to bind to primary antibodies that have been used to specifically recognize the target proteins, allowing for the detection of the target proteins through the fluorescent signal emitted by the dye-conjugated secondary antibodies.
Automatically generated - may contain errors
Lab products found in correlation
Image pro plus software, by Olympus (2 mentions)
Magnafire sp 2.1b software, by Olympus (1 mentions)
Rabbit anti caspase 3, by Santa Cruz Biotechnology (1 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Goat anti iba 1, by Santa Cruz Biotechnology (1 mentions)
Goat anti glial fibrillary acidic protein, by Santa Cruz Biotechnology (1 mentions)
Anti neun, by Abcam (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Ab15690, by Abcam (1 mentions)
Ab177487, by Abcam (1 mentions)
Ab7260, by Abcam (1 mentions)
Rabbit anti erbb4, by Abcam (1 mentions)
Mouse anti cd31, by Abcam (1 mentions)
Rabbit anti yap, by Cell Signaling Technology (1 mentions)
4 protocols using fluorescence dye conjugated secondary antibodies
1
Fluorescent Labeling of Caspase-3 and CHOP
2
Quantification of Neuronal Apoptosis
3
Immunofluorescence Staining of Brain Sections
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Immunofluorescence staining was performed as previously described [47 (link), 48 (link)]. Briefly, the animals were perfused with ice cold PBS and followed by 10% formalin under deep anesthesia. The animals' brains were further fixed in 10% formalin at 4°C for 48 h and dehydrated in 30% sucrose solution for a week. The brains were sectioned to 10 μm thick slices. The brain slices were stained overnight with following primary antibodies including rabbit anti-EP3 (1 : 50, 14357-1-AP, Proteintech, Rosemont, USA), mouse anti-NeuN (1 : 100, ab177487, Abcam, MA, USA), mouse anti-Iba1 (1 : 100, ab15690, Abcam, MA, USA), and mouse anti-GFAP (1 : 100, ab7260, Abcam, MA, USA) overnight at 4°C. On the second day, the slices were incubated with the respective fluorescence dye-conjugated secondary antibodies (1 : 200, Jackson ImmunoResearch, PA, USA) on dark condition for 2 h followed by DAPI staining. The staining was observed using a fluorescence microscope (Olympus, Melville, NY, USA).
4
Double-Immunofluorescence Staining of Cerebral Cortex Post-SAH
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Double-immunofluorescence staining of cerebral cortex was performed at 24 h after SAH as described previously (Xie et al., 2017 (link)). Sections were incubated overnight at 4°C with rabbit anti-ErbB4 (Abcam, Cambridge, MA, United States), rabbit anti-YAP (Cell Signaling Technology, Beverly, MA, United States), and mouse anti-CD31 (Abcam, Cambridge, MA, United States), followed by fluorescence dye-conjugated secondary antibodies (Jackson Immunoresearch, West Grove, PA, United States) for 3 h at 20°C. The sections were then visualized with a fluorescence microscope. Photomicrographs were analyzed using Image-Pro Plus software (Olympus, Melville, NY, United States).
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