Mice were anesthetized and immediately perfused with PBS and 4% paraformaldehyde (PFA) as previously described [32 (link)]. The brain was post-fixed in 4% PFA overnight and transferred to 30% sucrose in PBS. 50 μm thick sections were prepared by vibratome. Then, the brain sections were incubated with primary antibodies as following: anti-Caspase-1 (1:500, AdipoGen, CA, USA), anti-NeuN (1:300, Abcam, Cambridge, UK), anti-Lectin (1:300, DL-1177, Vector Laboratories, USA), anti-Nitrotyrosine (1:1000, Millipore, CA, USA) for overnight at 4 °C. After being rinsed three times, the sections were incubated with appropriate fluorescent secondary antibodies (Invitrogen, Carlsbad, CA) for 2 h at room temperature. Finally, the sections were stained with DAPI (0.5 μg/ml) for 5 min and mounted in Vectashield medium (Vector Laboratories, CA). Immunofluorescence images were performed with a Zeiss LSM800 confocal microscope (Carl Zeiss, Germany). The 3D surface visualization of grayscale intensity was plotted through ImageJ v1.50H and its “Interactive 3D Surface Plot” plug-in.
Anti lectin
Manufactured by Vector Laboratories
Sourced in United States
Anti-Lectin is a laboratory reagent used for the detection and quantification of lectins in various biological samples. Lectins are carbohydrate-binding proteins that play important roles in various biological processes. Anti-Lectin is a specific antibody that binds to lectins, allowing for their identification and measurement using techniques such as ELISA or immunoblotting.
Automatically generated - may contain errors
Lab products found in correlation
Lsm800 confocal microscope, by Zeiss (1 mentions)
Fluorescent secondary antibody, by Thermo Fisher Scientific (1 mentions)
Anti nitrotyrosine, by Merck Group (1 mentions)
Anti neun, by Abcam (1 mentions)
Vectashield medium, by Vector Laboratories (1 mentions)
Anti caspase 1, by Adipogen (1 mentions)
Hb 10104af, by Nikon (1 mentions)
Dxm1200, by Nikon (1 mentions)
Eclipse e600, by Nikon (1 mentions)
Anti akt polyclonal antibody, by Cell Signaling Technology (1 mentions)
Alexafluor 594 igg fraction monoclonal mouse anti biotin, by Jackson ImmunoResearch (1 mentions)
Anti brdu antibody, by Abcam (1 mentions)
Anti tubulin monoclonal antibody, by Abcam (1 mentions)
Growth factor reduced matrigel, by Corning (1 mentions)
3 protocols using anti lectin
1
Immunohistochemical Analysis of Mouse Brain
2
Whole-Mount Retina Immunohistochemistry
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Eyes were removed and fixed for 24 h with 4% PFA, at room temperature. Retinas were prepared as flattened whole mounts, by making four radial cuts, and labeled with anti-lectin (20 μg/mL) (Vector Laboratories, USA) and anti-phalloidin (1:100) (Biotium, USA) antibodies. Retinas were permeabilized by washing in PBST for 30 min, then, washed them in PBS 3 times for 10 min and, finally, anti-lectin was applied at 4 °C overnight. The following day, they were washed in PBS 3 times for 5 min and incubated in anti-phalloidin at 4 °C overnight. Retinas were mounted with an anti-fade reagent containing Hoechst-33258 to label nuclei (Molecular Probes, USA). Twenty micrometer-thick sagittal sections were assessed for potential anatomical alterations. Finally, whole-mount retinas and sagittal sections were imaged using a light microscope (× 400, Eclipse E600, Nikon Instruments Inc., Japan) equipped with a manually controlled specimen stage for X, Y, and Z axis, a color camera (model DXM1200, Nikon Instruments Inc., Japan), a fluorescent light source (HB-10104AF, Nikon Instruments Inc., Japan), and the image analysis software (NIS-Elements, Version 3.22, Nikon Instruments Inc., Japan).
3
Matrigel-based Protein Expression Assay
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Growth Factor Reduced Matrigel was purchased from Corning (356230, USA). Recombinant human ANXA2 protein was purchased from R&D System (D9409-AN-050, USA). Anti-BrdU antibody was purchased from Abcam (ab1893, USA). Anti-Lectin was purchased from Vectorlabs, (B-1415-2, USA). Anti-Tubulin monoclonal antibody was purchased from Abcam (ab210797, USA). Anti-A2R polyclonal antibody was purchased from Novus Biologicals (NBP-53077, USA). Anti-AKT polyclonal antibody was purchased from Cell Signaling Technology (9272s, USA). Anti-p-AKT polyclonal antibody was purchased from Cell Signaling Technology (4060s, USA). Anti-ERK monoclonal antibody was purchased from Santa Cruz Biotechnology (sc-81457, USA). Anti-phospho-ERK monoclonal antibody was purchased from R&D System (DYC1483-5, USA). Alexa Fluor® 488 A niPure donkey anti-goat IgG were purchased from Jackson ImmunoResearch Laboratories (705-545-147, USA). Alexa Fluor® 488 IgG fraction monoclonal mouse anti-Biotin were purchased from Jackson ImmunoResearch Laboratories (200-542-211, USA). Alexa Fluor® 594 IgG fraction monoclonal mouse anti-Biotin were purchased from Jackson ImmunoResearch Laboratories (200-582-211, USA).
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