bEnd.3 cells were washed thrice with PBS, and were permeabilized for 20 min. bEnd.3 cells were incubated with the primary antibodies for 16 h at 4 °C. The following primary antibodies were used: anti-rabbit AdipoR1 (1:500, Abcam), anti-goat AdipoR2 (1:500, Abcam), anti-rabbit NF-κB (1:500, Cell Signaling), anti-rabbit p-NF-κB (1:500, Cell Signaling), anti-rabbit CD31 (1:500, Abcam), and anti-rabbit Claudin 5 (1:500, Cell Signaling). After 16 h incubation, bEnd.3 cells were washed twice with PBS. bEnd.3 cells were incubated with each specific secondary antibody for 1 h and 30 min at room temperature. bEnd.3 cells were counterstained with 1 μg/ml 4', 6-diamidino-2-phenylindole (DAPI, 1:100, Invitrogen) for 10 min at room temperature. Images were obtained using confocal microscope (Carl Zeiss).
Rabbit anti cd31
Manufactured by Abcam
Sourced in United Kingdom, United States
Rabbit anti-CD31 is a primary antibody that recognizes the CD31 antigen, also known as platelet endothelial cell adhesion molecule (PECAM-1). CD31 is a transmembrane glycoprotein expressed on the surface of endothelial cells and is involved in cell-cell adhesion. This antibody can be used to detect and localize CD31 in various applications, such as immunohistochemistry and flow cytometry.
Automatically generated - may contain errors
Lab products found in correlation
Mouse anti α sma, by Abcam (5 mentions)
Ix81 microscope, by Olympus (4 mentions)
Rat anti ki67, by Thermo Fisher Scientific (3 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Rabbit anti ki67, by Abcam (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (2 mentions)
Alexa fluor 647 goat anti rabbit, by Thermo Fisher Scientific (2 mentions)
Ix71 fluorescence microscope, by Olympus (2 mentions)
Viobright 515, by Miltenyi Biotec (2 mentions)
Cy3 conjugated mouse anti sma antibodies, by Merck Group (2 mentions)
Rabbit anti occludin, by Thermo Fisher Scientific (2 mentions)
Rabbit anti α smooth muscle actin, by Abcam (2 mentions)
Alexa fluor 488 conjugated goat anti mouse secondary antibody, by Abcam (2 mentions)
Rabbit anti α sma, by Abcam (2 mentions)
Rabbit anti nf κb, by Cell Signaling Technology (1 mentions)
Confocal microscope, by Zeiss (1 mentions)
Anti mouse nk1, by BioLegend (1 mentions)
Vectashield hardset mounting medium with dapi, by Vector Laboratories (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Axioskop 2 mot plus, by Zeiss (1 mentions)
57 protocols using rabbit anti cd31
1
Immunofluorescence Analysis of bEnd.3 Cells
2
Quantifying Tumor Microenvironment Characteristics
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Specimens of the harvested livers and tumor tissues were fixed in 4 % formalin followed by embedding in paraffin. Tissue sections (4 μm) were subjected to histopathology (H&E staining), immunohistochemical (IHC) staining and in situ apoptosis detection by TUNEL assay. For IHC, after dewaxing, antigen retrieval and endogenous peroxidase blocking steps the sections were subjected to staining procedures with purified monoclonal anti-mouse CD11b (BD PharMingen, San Jose, CA); anti-mouse NK1.1 (Biolegend, San Diego, CA); and anti-rabbit CD31 (Abcam, Cambrige, MA) primary antibodies, respectively. Subsequently, biotinylated goat anti-mouse or anti-rabbit immunoglobulins as secondary antibodies and streptavidin peroxidase complex reagent were applied. The visualization signals were developed with diaminobenzidine (DAB) chromogen substrate (DAKO, Carpinteria, CA and the slides were counterstained with Meyer’s hematoxylin and dehydrated through a series of ethanol and xylenes. In addition, intratumor microvessel density was microscopically counted in absolute values as previously described [29 (link), 34 (link)]. Finally, for detection of in situ apoptosis in the tumor tissues, TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick-end labeling) assay was performed according to the manufacturer’s protocol (Roche Molecular Biochemicals) as described previously [34 (link)].
3
Immunofluorescence Analysis of CD31 in Cells
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Immunofluorescence was performed on cells grown in matrigel. The cells were fixed with 4% paraformaldehyde (Sigma-Aldrich, Missouri) for 20 minutes and permeabilized with 1:1 methanol and acetone for 15 minutes in 4C°. After blocking in 5% goat serum, cells were probed with anti-rabbit CD31 (1:100; abcam). Alexa Fluor goat anti-rabbit IgG (1:500, Carlsbad, CA) was used as secondary antibody. The cells in the matrigel slides were mounted with cover slips using Vectashield hard set mounting medium with DAPI (Vector Laboratories, Burlingame, CA) and the immunofluorescence images were visualized using an inverted fluorescence microscope (Axioskop 2 MOT plus, Zeiss) and Axiovision software.
4
Characterization of Testisin Variants
5
Protein Extraction and Western Blot Analysis
6
Comprehensive Cardiac Tissue Analysis
7
Immunofluorescent Staining of Tumor Tissues
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IHC staining of the tumors was performed as described [27 (link)]. Paraffin-embedded sections were stained ON at 4°C with a rabbit anti-CD31 (4 μg/ml, Abcam) and a rat anti-Ki67 (1 μg/ml, eBioscience) antibody. After washing, sections were incubated for 3 h at RT with Alexa Fluor 568 goat anti-rabbit antibody (4 μg/ml; Molecular probes) and Dylight 650 conjugated goat anti-rat antibody (4 μg/ml; Thermofisher) and examined by fluorescence microscopy.
8
Culturing Human Bronchial Epithelial Cells
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Human bronchial epithelial (16HBE) cells were obtained from Shanghai Institute for Biological Science. Cells were cultured in RPMI 1640 (HyClone, UT, USA) supplemented with 100 U/mL penicillin, 100 mg/mL streptomycin, and 10% heat-inactivated fetal bovine serum (BIO International, Auckland, New Zealand). All cells were maintained at 37 °C in a humidified incubator with 5% carbon dioxide. Human recombinant CCN1 proteins were obtained from PeproTech (Shanghai, China). The rabbit anti-EpCAM as well as rabbit anti-CD31 were purchased from Abcam (HK, China). Mouse anti-CCN1 and horseradish peroxidase-coupled secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA).
9
Quantitative Analysis of Tumor Angiogenesis
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After tumor removal, the fresh tumors were immediately embedded in optimum cutting temperature compound (Sakura, Zoeterwoude, Netherlands) and sectioned (5 mm), followed by staining with hematoxylin and eosin (H&E), CD31 antibody, and oil red O. Briefly, sections mounted on slides were dehydrated in ethanol, rinsed in PBS containing Tween 20 (PBST), and incubated with 0.3% hydrogen peroxide for 15 min. After washing with PBST, sections were blocked by incubation in 3% bovine serum albumin for 30 min, followed by overnight incubation with primary antibody (rabbit anti-CD31 diluted 1 : 50; Abcam). Slides were washed with PBST followed by a 1 h incubation with HRP-conjugated goat anti-rabbit secondary antibody (1 : 4000, Abcam), rinsed in PBST, and exposed to 3,3′-diaminobenzidine (Solarbio, Beijing, China). Then, counterstaining was performed with hematoxylin (Solarbio, Beijing, China). For H&E staining, sections were stained in hematoxylin for 3 min, washed in water, and then exposed for 5 min to eosin (Solarbio). The immunostaining results were analyzed using imaging software (Olympus, Tokyo, Japan).
10
Immunohistochemical Analysis of Wound Healing
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Histological sections (4 µm) from paraffin-embedded fixed tissues were stained with haematoxylin-eosin (H&E) for microscopic analysis of wound healing. Moreover, sections were subjected to immunohistochemistry following antigen retrieval as described previously33 (link). Tissue sections were blocked in 3% blocking serum for 30 min at room temperature, incubated with rabbit anti-human collagen type I (5 µg/mL, Rockland Immunochemicals, Pennsylvania, USA), rabbit anti-human collagen type III (5 µg/mL, Rockland Immunochemicals, Pennsylvania, USA), rat neutrophil marker (NIMP-R14, 0.5 µg/mL, Santa Cruz Biotechnology, Texas, USA) and rabbit anti-CD31 (0.2 µg/mL, Abcam, Cambridge, UK), F4/80 (5 µg/mL, Bio-Rad, NSW, Australia), YM-1 (0.37 µg/mL, StemCell Technologies, Vancouver, Canada), actin smooth muscle antibody (αSMA, 0.7 µg Agilent Technologies, VIC, Australia) antibodies overnight at 4 °C. Subsequently, tissue sections were stained with secondary goat anti-rabbit IgG, Alexa Fluor 488, goat anti-rabbit IgG, Alexa Fluor 568 or goat anti-rat IgG Alexa Fluor 488 (5 µg/mL, Invitrogen, CA, USA). To visualize nuclei and overall tissue architecture, sections were stained for 5 min in 4′,6-diamidino-2-phenylindole (DAPI, 1:5000 of 1 mg/mL stock). Sections were imaged with Olympus IX81 microscope (Olympus, Tokyo, Japan).
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