The sections were fixed with 4% paraformaldehyde for 30 min, permeabilized in PBS containing 1% Triton X-100 for 10 min, followed by washing three times with PBS, and blocked with 10% goat serum in PBS for 30 min. Then, the sections were incubated with primary antibodies against CD31 (abcam, UK) or collagen type II (abmart, China) in the Superblock solution overnight at 4 °C. On the next day, sections were washed with PBS three times for 5 min each, followed by incubation with the Goat Anti-Rabbit IgG (abcam, UK) or Goat Anti-Rat IgG H&L (abcam, UK) for 2 h at room temperature. Nuclei were counterstained with 4,6-diamidino-2-phenylindole (DAPI) for 2 min. After washing, the tissue was observed under the CLSM in the darkroom. Quantification for CD31 and COL II intensities were determined based on the obtained mHIC images via ImageJ software.
Primary antibodies against cd31
Manufactured by Abcam
Sourced in United Kingdom
Primary antibodies against CD31 are laboratory reagents used to detect and study the CD31 protein, also known as platelet endothelial cell adhesion molecule (PECAM-1). CD31 is a cell surface glycoprotein expressed on the surface of endothelial cells and some immune cells. These antibodies can be used in various research applications, such as immunohistochemistry, flow cytometry, and Western blotting, to identify and characterize CD31-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Goat anti rat igg h l, by Abcam (1 mentions)
Goat anti rabbit igg, by Abcam (1 mentions)
Antigen retrieval solution, by Agilent Technologies (1 mentions)
3 3 diaminobenzidine dab substrate, by Vector Laboratories (1 mentions)
Vegfa, by Abcam (1 mentions)
Sp5 2 confocal uorescent microscope, by Leica (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Vector Laboratories (1 mentions)
5 protocols using primary antibodies against cd31
1
Immunofluorescent Analysis of Endothelial and Cartilage Markers
2
CD31 Immunohistochemical Staining of Formalin-Fixed, Paraffin-Embedded Tissues
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Fresh tumours were fixed with 10% neutral formalin for 3 day and then embedded in paraffin. The tissues were sectioned at the thickness of 4 μm. Immunohistochemistry staining for CD31 was performed using the Histostain‐Plus Kit (Kangwei). After sequential treatments, tissue sections were incubated with normal serum for 30 minutes, then incubated with the control IgG and primary antibodies against CD31 (Abcam, 1:200) at 4℃ overnight and then HRP‐labelled secondary antibody for 30 minutes. The 3,3‐diaminobenzidine (DAB) solution was used for development of brown colour to identify the expression of CD31.
3
Immunohistochemical Quantification of CD31 in Mouse Tumors
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The immunohistochemical analysis was perfomed as previously described.54 (link) Briefly, unstained sections of mouse tumor tissue were deparaffinized and rehydrated. Antigens were retrieved by using Dako antigen retrieval solution. Endogenous peroxidase was blocked with 3% hydrogen peroxide. Primary antibodies against CD31 (Abcam) were incubated overnight at 4°C. The next day, a goat anti-rabbit horseradish peroxidase secondary antibody (Jackson ImmunoResearch Laboratories), diluted in blocking solution, was added and the samples were incubated for 1 hr at room temperature. Slides were developed with 3,3′-diaminobenzidine (DAB) substrate (Vector Laboratories) and counterstained with Gill no. 3 hematoxylin solution. To quantify CD31 expression, the positive (DAB-stained) cells were counted in five random fields per slide.
4
Regenerated Tibia Histological Analysis
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Regenerated tibia samples (n = 3 per group at each time termination) were harvested 2, 4, 6, and 8 weeks after scaffold implantation and submersion fixed in 4% PFA for 3 days. The samples were then dehydrated in graded concentrations of alcohols, and then decalcified with 10% EDTA for 6 weeks. The samples were then embedded in paraffin and sectioned into 5-μm-thick sections along the long axis in the midsagittal plane. For histological analyses, sections were stained with hematoxylin-eosin (HE) and Masson's trichrome stains. For immunohistochemical (IHC) staining, the sections were incubated in primary antibodies against CD31, CD34, VEGF-A, or osteocalcin (OCN) (1:200; Abcam, Cambridge, UK) overnight at 4 °C, followed by species-appropriate HRP-conjugated secondary antibodies (1:1000, Jackson Research). Immunoreactivity was visualized with 3,3′-diaminobenzidine (DAB). Integrated optical density (IOD) was measured in five randomly selected visual fields per section using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
5
Quantifying Muscle Angiogenesis in Ischemic Limbs
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After 28 days of ischemia, mice were euthanized by intraperitoneal administration of ketarom (10 mg/kg xylazine mixed with 100 mg/kg ketamine, 0.2 mL/ 20 g) and blood samples were obtained. Subsequently, ischemic limb muscle tissue was harvested and embedded in OCT (Sakura, Japan). Cross-sections (6 µm) of muscle were subjected to CD31 histological immunostaining. Brie y, sections were blocked with 10% donkey serum at 37°C for 1 h. Primary antibodies against CD31 (Abcam, UK), an endothelial cell marker, were applied at 4°C overnight, and subsequently, Alexa 488 uor-conjugated secondary antibodies (Abcam, UK) were used. Cell nuclei were counterstained with DAPI (Vector Laboratories, USA). CD31-positive cells were observed and quanti ed under a Leica SP5 II confocal uorescent microscope.
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