Frozen coronal sections (7 μm) were collected near the collagenase injection site and washed three times in PBS at room temperature. Antigen retrieval was performed by placing slides in a pressure cooker containing citrate buffer (pH 6.0) for 4 min. Tissues were permeabilized by 1% triton-X100 in PBS for 30 min, washed in PBS, and blocked in 1% BSA in PBS for 1 hr. Slides were incubated overnight at 4°C in 1% BSA containing the following primary antibodies: mouse anti-3-NT (1:500, Sigma), rabbit anti-iNOS (1:300, Cell Signaling), rabbit anti-GFAP (1:1000, Abcam), mouse anti-Iba1 (1:1000, Wako), rabbit anti-CD31 (1:300, EMD Millipore), and mouse anti-NeuN (1:1000, Sigma). The slides were washed in PBS, and then incubated for 1 hr at room temperature in 1% BSA containing Alexa Fluor®594-conjugated goat anti-rabbit and DyLight™488-conjugated goat anti-mouse (1:500, Jackson ImmunoResearch) secondary antibodies. After a final PBS wash, slides were mounted with SlowFade® Diamond Antifade Mountant with DAPI (ThermoFisher) and a coverslip. Images were captured using fluorescent microscopy. Mean fluorescence intensity and cell count per field were quantified by a person who was blinded to the experimental groups, using NIH image J software.
Rabbit anti cd31
Manufactured by Merck Group
Sourced in United States
Rabbit anti-CD31 is a laboratory reagent used for the detection and characterization of the CD31 protein, also known as Platelet Endothelial Cell Adhesion Molecule 1 (PECAM-1). CD31 is a cell surface glycoprotein expressed on the surface of endothelial cells, platelets, and certain immune cells. This product can be used in various immunological and cell biology applications, such as flow cytometry, immunohistochemistry, and Western blotting.
Automatically generated - may contain errors
Lab products found in correlation
α smooth muscle actin, by Abcam (2 mentions)
α sma, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (2 mentions)
Mouse anti neun, by Merck Group (1 mentions)
Mouse anti iba 1, by Fujifilm (1 mentions)
Slowfade diamond antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 594 conjugated goat anti rabbit, by Jackson ImmunoResearch (1 mentions)
Rabbit anti inos, by Cell Signaling Technology (1 mentions)
Rabbit anti gfap, by Abcam (1 mentions)
Axioskop 2 mot plus, by Zeiss (1 mentions)
Axiovision le software, by Zeiss (1 mentions)
Vectashield hardset mounting medium, by Vector Laboratories (1 mentions)
4 protocols using rabbit anti cd31
1
Immunohistochemical Analysis of Spinal Cord Injury
2
CD31 Expression Analysis in Cell Lines
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Immunofluorescence analysis was performed on HUVEC, M1102-PAX2 and SKOV3 cells grown in Matrigel. The cells were fixed with 4% paraformaldehyde (Sigma-Aldrich; Merck KGaA) for 20 min and permeabilized with 1:1 methanol and acetone for 15 min, both at 4˚C. After blocking for 1 h at room temperature in 5% goat serum (Sigma-Aldrich; Merck KGaA), cells were probed with rabbit anti-CD31 (1:100; cat. no. ab222783; Abcam) for 1 h at room temperature. Alexa Fluor goat anti-rabbit IgG (1:500, cat. no. A32731; Thermo Fisher Scientific, Inc.) was used as the secondary antibody for 1 h at room temperature. The cells were mounted on Matrigel slides with VectaShield hard set mounting medium containing DAPI (Vector Laboratories, Inc.), and immunofluorescence was visualized at 100X magnification using an inverted fluorescence microscope (Axioskop 2 MOT plus) and AxioVision LE software (version 4.8.2; Carl Zeiss AG).
3
Immunohistochemical Quantification of Vascular Structures
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Tissues were rapidly excised and fixed in neutral buffered formalin, embedded in paraffin, and 5 μm sections were stained with H&E. Immunohistochemistry was performed to determine capillaries and small arteries using primary antibodies as follows: rabbit anti-CD31 (Chemicon, Temecula, CA, USA) and α-smooth muscle actin (α-SMA, Epitomics; Burlingame, CA, USA), then incubated with respective secondary antibodies and nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, Vector Laboratories, Burlingame, CA, USA). The number of positive staining of each section was counted in five different fields (original magnification, ×400).
4
Immunohistochemical Quantification of Vascular Structures
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