Free floating brain sections (25 μm) from APOE3 and APOE4 mice were blocked for 1 h in a PBS solution containing 5% horse serum (Invitrogen, Carlsbad, CA) and 0.4% Triton X-100. Sections were then incubated over-night at 4°C with primary antibodies in the blocking solution: biotinylated Lectin antibody (Vector laboratories, Burlingame, CA) and rabbit anti-insulin receptor (INSR, 1:100; Fitzgerald; Acton, MA). After incubation with primary antibodies, slices were exposed to Alexa Fluor-488 conjugated streptavidin (Invitrogen, Carlsbad, CA) or Alexa Fluor-647 conjugated donkey anti-rabbit secondary antibodies (1:1000; Invitrogen, Carlsbad, CA). Then, slices were counterstained with 4’, 6-diamino-2-phenylindole (DAPI; Invitrogen) for 10 min, mounted on SuperFrost Plus slides and treated with 0.5% Sudan black (in 70% methanol) for 5 min. Finally, slides were placed under coverslips with Mowiol mounting media. Immunofluorescence was examined using an epifluorescence microscope (Olympus Provis AX70; Olympus, Melville, NY) and photographs were taken using a Spot digital camera (Diagnostic Instruments, Sterling Heights, MI). All images were prepared for illustration with Fiji/ImageJ software.
Biotinylated lectin antibody
Manufactured by Vector Laboratories
Sourced in United States
The Biotinylated Lectin antibody is a laboratory product designed to detect the presence of lectin, a type of carbohydrate-binding protein, in biological samples. The antibody is tagged with biotin, a small molecule that can be used to amplify or visualize the signal from the lectin detection.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 conjugated streptavidin, by Thermo Fisher Scientific (1 mentions)
4 6 diamino 2 phenylindole dapi, by Thermo Fisher Scientific (1 mentions)
Provis ax70, by Olympus (1 mentions)
Horse serum, by Thermo Fisher Scientific (1 mentions)
3 3 diaminobenzidine (dab), by Thermo Fisher Scientific (1 mentions)
Ds ri1 camera, by Nikon (1 mentions)
Bx51 microscope, by Nikon (1 mentions)
2 protocols using biotinylated lectin antibody
1
Brain Section Immunofluorescence in APOE Mice
2
Lectin-Based Myocardial Capillary Quantification
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Glycoprotein lectin was used to identify capillary endothelial cells in all myocardial sections. Sections were dewaxed, rehydrated and antigen retrieval carried out as above. Slides were blocked with 0.3% H2O2, followed by 6% goat serum. Sections were stained with Biotinylated Lectin Antibody (B-1105, Vector Laboratories, Burlingame, CA, USA) in a 1:300 dilution. Following overnight incubation, mouse Avidin-Biotin Complex (ABC; Thermo Scientific) was added to the stained sections. The chromogen used for visualization of the lectin signals was 3,3′-Diaminobenzidine (DAB), made using DAB metal concentrate (10x in stable peroxide substrate buffer (1x) at a 1:100 dilution (Thermo Scientific). Stains were imaged using an Olympus BX51 microscope and Nikon DS-Ri1 camera with NIS elements software. Microvessel density, measured as small vessel counts per tissue area (µm2), was calculated using an automated analysis program created on NIS Analysis elements software.
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