Anti mouse cd31 monoclonal antibody

Manufactured by Abcam

Sourced in United Kingdom, United States

The Anti-mouse CD31 monoclonal antibody is a laboratory reagent used to detect the presence of the CD31 protein, also known as platelet endothelial cell adhesion molecule (PECAM-1), in mouse samples. CD31 is a transmembrane glycoprotein that plays a role in cell-cell adhesion and is commonly used as a marker for endothelial cells.

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Lab products found in correlation

Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) Confocal microscope, by Nikon (1 mentions) Goat anti rabbit igg fitc, by Boster Bio (1 mentions) Bx50 fluorescence microscope, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Rabbit anti-mouse CD31 monoclonal antibody Rat anti-mouse CD31 monoclonal antibody CD31 mouse monoclonal antibody Anti-CD31 monoclonal antibody Anti-mouse CD31 antibody Mouse anti-CD31 Anti-CD31 antibody Monoclonal mouse antibody CD31 antibody Anti-CD31

7 protocols using anti mouse cd31 monoclonal antibody

Quantifying VEGF and HIF-1α Expression in Tumors

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The sections were stained for VEGF and HIF-1α expression as well as microvessel density (MVD) utilizing the Catalyzed Signal Amplification System (Dako Denmark A/S, Glostrup, Denmark). After deparaffinization and rehydration of the slides, the process of antigen retrieval (Dako Denmark A/S) lasted for 45 minutes at 97°C. Hematoxylin was used to counterstain the nuclei. Primary antibodies used in the experiments were as follows: mouse anti-CD31 monoclonal antibody 1:300 (Abcam, Hong Kong, People’s Republic of China), mouse anti-VEGFα monoclonal antibody (1:1,500) (Abcam), and rabbit anti-HIF-1α polyclonal antibody (1:500) (Sigma-Aldrich Co., St Louis, MO, USA).
For each slide, two or three fields were counted (400× magnification) to access the frequency of VEGF-positive cytoplasms and HIF-1α-positive nuclei. Twenty-two fields were counted in each group. VEGF and HIF-1α expression were determined by the means of calculating the percentage of immunoreactive cells among the population of tumor cells. The MVD was determined to be the average microvessels in each field (400× magnification).22 (link) Any area of obvious CD31-positive staining was considered as a single vessel. Twenty-four fields were checked in ten different tumors in every group. At least two of the authors, who were blinded to the identity of the sections, analyzed the section using a light microscope.

Chen C.S., Zhao Q., Qian S., Li H.L., Guo C.Y., Zhang W., Yan Z.P., Liu R, & Wang J.H. (2015). Ultrasound-guided RNA interference targeting HIF-1 alpha improves the effects of transarterial chemoembolization in rat liver tumors. OncoTargets and therapy, 8, 3539-3548.

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Quantifying Angiogenesis in Rabbit Capsule

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After general anesthesia, the capsule tissue of each rabbit was fully exposed. Then, the capsule tissues were harvested and placed in liquid nitrogen or 4% paraformaldehyde. The samples were embedded in paraffin and sectioned into 4-μm thick sections, and hematoxylin and eosin (H&E) staining was performed according to the standard protocols. In addition, the sections were deparaffinized and blocked with 3% bovine serum albumin (BSA) for 30 min followed by incubation with a mouse anti-CD31 monoclonal antibody (1 : 500; Abcam, Cambridge, UK). The images were acquired using fluorescence microscopy. Each slide was carefully examined at ×40 magnification to identify the area with the highest density of vessels. Then, 3 fields of view were selected at ×200 magnification. The number of CD31-positive vessels and the area ratios were quantified using ImageJ software (National Institutes of Health, Bethesda, USA).

Yu M., Bian Y., Wang L, & Chen F. (2021). Low-intensity pulsed ultrasound enhances angiogenesis in rabbit capsule tissue that acts as a novel vascular bed in vivo. Advances in clinical and experimental medicine : official organ Wroclaw Medical University, 30(6).

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Quantifying Tumor Angiogenesis via CD31 Staining

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At the end of the protocol, mice were euthanized and perfused with PBS. Tumors were fixed with formalin, paraffin-embedded and sectioned (5μm). Endothelial cells were stained with an anti-mouse CD31 monoclonal antibody (1:50, Abcam, Cambridge, MA). For quantitation of vessel density, the areas of highest neovascularization, “hot spots”, were identified by scanning the sections at low power (200X) followed by vessel counts at high power (400X), as previously described [10 (link)]. For each tumor section, five hot spots were identified and vessels were counted.

Wang J., Tran J., Wang H., Luo W., Guo C., Harro D., Campbell A.D, & Eitzman D.T. (2015). Melanoma tumor growth is accelerated in a mouse model of sickle cell disease. Experimental Hematology & Oncology, 4, 19.

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Immunostaining of CD31 in Skin Sections

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For detection of CD31 expression, 5-μm-thick paraffin-embedded sections of skin were deparaffinized and rehydrated. For permeabilization, the samples were incubated with a 0.05% Triton X-100 solution at room temperature for 10 min and blocked with a 5% NGS for 1 h. Then, the cells were stained with anti-mouse CD31 monoclonal antibody (Abcam) overnight at 4 °C and secondary antibody labeled with Alexa Fluor 488 (Molecular Probes) for 1 h. Before imaging, nuclei were counterstained with DAPI. The images were captured by a confocal microscope (Nikon).

Lee B.C., Kim J.J., Lee J.Y., Kang I., Shin N., Lee S.E., Choi S.W., Cho J.Y., Kim H.S, & Kang K.S. (2019). Disease-specific primed human adult stem cells effectively ameliorate experimental atopic dermatitis in mice. Theranostics, 9(12), 3608-3621.

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Quantifying Tumor Angiogenesis via CD31 Immunostaining

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Cryosections (30 um) of the removed plugs were obtained. Plugs were then immunostained with an anti-mouse CD31 monoclonal antibody (1:300, Abcam, Cambridge, MA, UK) and a secondary goat anti-rabbit Alexa 594 antibody (1:5000, Life Technologies, Carlsbad, CA, USA), according to the manufacturer’s protocol.
The immunofluorescence images were analyzed using NIH-Image J software (National Institutes of Health, Bethesda, Maryland, USA). A free-hand ROI was drawn around the tumor, and CD31-positive cells were selected using the global histogram-derived thresholding method [19 (link)]. The percentage area of CD31-positive cells was calculated by dividing the area of the CD31-positive cells by the area of the tumor ROI.

Kim J., Choi S.H., Ham S.J., Cho Y.C., Lee S.I., Kang J., Woo D.C., Lee W.S., Yoo J.S., Kim K.W, & Choi Y. (2018). Evaluation of drug mechanism and efficacy of a novel anti-angiogenic agent, TTAC-0001, using multi-modality bioimaging in a mouse breast cancer orthotopic model. PLoS ONE, 13(1), e0187063.

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Immunofluorescence Staining of Angiogenesis Markers

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Five rats from each group were chosen at each time point. Normal saline and 4% paraformaldehyde were perfused through the heart to stabilize tissues, which were cut into frozen slices (40-µm thick) through the coronal plane of the caudate putamen. Their membranes were ruptured with 0.3% Triton and the sections were sealed with normal animal serum. Drops of primary antibody were added after the slices were rinsed. Primary antibody concentrations: anti-CD31 mouse monoclonal antibody (1:500, incubated for 48 hours at 4°C; Abcam, Cambridge, MA, USA), anti-Ang-l rabbit polyclonal antibody (1:100, incubated for 48 hours at 4°C; Abcam), and anti-Ang-2 rabbit polyclonal antibody (1:500, incubated for 48 hours at 4°C; Abcam). Appropriate fluorescent secondary antibodies were added after rinsing with PBS: goat anti-mouse IgG-Cy3 (1:50, incubated for 2 hours at 20°C; Boster, Wuhan, China) and goat anti-rabbit IgG-FITC (1:50, incubated for 2 hours at 20°C; Boster). The nucleus was marked after 5-minute 4′,6-diamidino-2-phenylindole (DAPI) staining. Slices were observed and photographed under a BX50 fluorescence microscope (Olympus Co., Tokyo, Japan). A confocal microscope (Zess, Oberkochen, Germany) was used to observe the stained slices and scanned images.

Xiao H., Huang Q., Wang J.Q., Deng Q.Q, & Gu W.P. (2016). Effect of ephrin-B2 on the expressions of angiopoietin-1 and -2 after focal cerebral ischemia/reperfusion. Neural Regeneration Research, 11(11), 1784-1789.

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Evaluation of H-MnO2-PEG/TP Tumor Targeting

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CAL27 tumor-bearing mice were injected intravenously with PBS or H-MnO₂-PEG/TP. Liver, kidney, and lungs bearing tumors were surgically excised 20- or 120-min post injection. Tissue sections (4 mm) were stained with hematoxylin and eosin (HE). Terminal deoxynucleotide transferase dUTP notch end labeling (TUNEL) was used to detect apoptotic cells. To detect oxidation, the tumor sections were treated with mouse anti-HIF1α primary antibody (dilution 1:200, Abcam Inc. USA) and Alexa Flour^® 488-conjugated goat anti-rabbit secondary antibody (dilution 1:200, CST Inc. USA) following the instructions. Tumor blood vessels were stained by anti-CD31 mouse monoclonal antibody (dilution 1:200, Abcam Inc.) and Alexa Flour^® 555-conjugated goat anti-mouse secondary antibody (dilution 1:200, CST Inc., USA), subsequently. Cell nuclei were stained with DAPI (dilution 1:5000, Invitrogen, USA). The obtained slices were observed by confocal microscopy (Leica SP5, Germany).

Zhou Z.H., Liang S.Y., Zhao T.C., Chen X.Z., Cao X.K., Qi M., Huang Y.Y., Ju W.T., Yang M., Zhu D.W., Pang Y.C, & Zhong L.P. (2021). Overcoming chemotherapy resistance using pH-sensitive hollow MnO2 nanoshells that target the hypoxic tumor microenvironment of metastasized oral squamous cell carcinoma. Journal of Nanobiotechnology, 19, 157.

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