Anti cd31 antibody

Manufactured by Thermo Fisher Scientific

Sourced in United States

The Anti-CD31 antibody is a laboratory reagent used to detect the presence of CD31, also known as PECAM-1, a cell surface marker expressed on endothelial cells. This antibody can be utilized in various immunoassay techniques, such as flow cytometry, immunohistochemistry, and Western blotting, to identify and quantify cells expressing CD31.

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Lab products found in correlation

Mouse endothelial culture medium, by Cell Biologics (2 mentions) Dynabead, by Thermo Fisher Scientific (2 mentions) Collagenase 1, by Worthington (2 mentions) Facsaria 2, by BD (2 mentions) Leica vt1000 s vibrating blade microtome, by Leica Microsystems (1 mentions) Alexa fluor 555 dye, by Thermo Fisher Scientific (1 mentions) Anti cd68 antibody, by Bio-Rad (1 mentions) Alexa fluor 488 dye, by Thermo Fisher Scientific (1 mentions) Eclipse e400, by Nikon (1 mentions) Anti ve cadherin, by Abcam (1 mentions) Anti ki 67 antibody, by Merck Group (1 mentions) Anti cd68 antibody, by Abcam (1 mentions) Anti β actin, by Merck Group (1 mentions) Anti zo 1 antibody, by Santa Cruz Biotechnology (1 mentions) Facscalibur flow cytometer, by BD (1 mentions) Cycloheximide (chx), by Santa Cruz Biotechnology (1 mentions) Anti vegfr 1, by Santa Cruz Biotechnology (1 mentions) Anti fgfr 1 antibody, by Santa Cruz Biotechnology (1 mentions) Anti vcam 1, by Thermo Fisher Scientific (1 mentions) Anti icam 1, by Thermo Fisher Scientific (1 mentions)

15 protocols using anti cd31 antibody

Imaging of Th9 Cell Migration in Lungs

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5×10⁶ TdTomato Th9 cells were injected i.v. into WT and CIKS^−/− or CCL17^+/− and CCL17^−/− GFP-reporter mice. 24h later mice were exposed to rIL-25 i.n. (500ng/mouse/day, Biolegend, San Diego CA) for two consecutive days. 24h later mice were given a lethal overdose of avertin and exsanguinated. Lung sections were prepared for live cell imaging as described (42 (link)). Briefly, 1.5% of low-melt agarose inflated tissues were sliced into 300–350 μm sections using Leica VT1000-S-Vibrating-Blade-Microtome (Leica Microsystems, Richmond Hill ON, Canada) on ice-cold PBS. Tissue sections were stained with fluorescently labeled anti-CD31 antibody (eBioscience, San Diego, CA) for 2h on ice. Tissues were imaged using Leica DMi8-inverted-5-channel confocal microscope equipped with an Environmental Chamber (NIH Division of Scientific Equipment and Instrumentation Services) to maintain 37°C and 5% CO₂. Post-acquisition mages were processed using Imaris (Bitplane, Concord MA).

Claudio E., Wang H., Kamenyeva O., Tang W., Ha H.L, & Siebenlist U. (2019). IL-25 orchestrates activation of Thelper cells via conventional dendritic cells in tissue to exacerbate chronic HDM-induced asthma pathology. Journal of immunology (Baltimore, Md. : 1950), 203(8), 2319-2327.

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Multiplex Immunofluorescence for Tumor Microenvironment

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Frozen tissue sections of tumor and normal tissues were used for single or double immunofluorescence labeling. An antibody specific for human IGF1R with a cross reactivity for mouse IGF1R was obtained from Sigma-Aldrich (dilution 1:200). An anti-human fibroblast active protein antibody (Santa Cruz Biotechnology, dilution 1:100) was used for identification of active human fibroblasts in tumor stroma. Anti CD68 antibody (AbD Serotec, dilution 1:100) and CD163 antibody (Santa Cruz Biotechnology, dilution 1:200) were used to identify macrophages, and anti-CD31 antibody (eBioscience, dilution 1:200) was for labeling endothelial cells in tumor blood vessels. Mouse monoclonal anti-CK19 antibody was from Sigma-Aldrich (dilution 1:100). Alexa Fluor 488 dye (green, Invitrogen, dilution 1:500) or Alexa Fluor 555 dye (red, Invitrogen, dilution 1:500) labeled secondary antibodies were used to detect biomarker-positive cells. Images were taken using fluorescence microscopy (Nikon Eclipse E400, Tokyo, Japan).

Zhou H., Qian W., Uckun F.M., Wang L., Wang Y.A., Chen H., Kooby D., Yu Q., Lipowska M., Staley C.A., Mao H, & Yang L. (2015). IGF1 Receptor Targeted Theranostic Nanoparticles for Targeted and Image-Guided Therapy of Pancreatic Cancer. ACS nano, 9(8), 7976-7991.

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Antibody Sourcing for Cell Analysis

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The anti-CD31 antibody was obtained from eBioscience (San Diego, CA, UA). The anti-VE-cadherin and anti-CD68 antibody were obtained from Abcam (Cambridge, MA, USA). The anti-Ki67 antibody was purchased from Merck Millipore (Burlington, MA, USA). Anti-β-actin was obtained from Millipore Sigma (Burlington, MA, USA). The antibody against mouse Angpt1 antibody was from Gene Tex Inc. (San Antonio, Tx, USA).

Chiang W.C., Huang Y.C., Fu T.I., Chen P.M., Chang F.C., Lai C.F., Wu V.C., Lin S.L, & Chen Y.M. (2019). Angiopoietin 1 influences ischemic reperfusion renal injury via modulating endothelium survival and regeneration. Molecular Medicine, 25, 5.

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Identification of Hippocampal Microvascular Endothelial Cells

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CD31ZO-1 double positive hippocampus microvascular endothelial cells were identified using an anti-CD31 antibody (eBioscience, San Diego, USA) and an anti-ZO1 antibody (Santa Cruz Biotechnology). Cell surface staining of freshly isolated brain single cell suspensions was performed with the fluorescently labelled antibodies at 4°C for 30 minutes. All flow cytometry measurements were carried out using a FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA, USA). Analysis was performed using CellQuestTM software, version 3.1 (Becton Dickinson).

Casciati A., Dobos K., Antonelli F., Benedek A., Kempf S.J., Bellés M., Balogh A., Tanori M., Heredia L., Atkinson M.J., von Toerne C., Azimzadeh O., Saran A., Sáfrány G., Benotmane M.A., Linares-Vidal M.V., Tapio S., Lumniczky K, & Pazzaglia S. (2016). Age-related effects of X-ray irradiation on mouse hippocampus. Oncotarget, 7(19), 28040-28058.

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Evaluation of Anti-Angiogenic Drug Effects

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Sunitinib and sorafenib were from LC Laboratories (Woburn, MA, USA). Sodium bicarbonate and HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) were from Sigma-Aldrich. Recombinant human VEGF was from Peprotech (#100-20-10). Cycloheximide was purchased from Santa Cruz biotechnology (#SC-3508). For Western blot analysis, the following primary antibodies and concentrations were used: anti-phospho-Akt antibody (1:500) (#4060; Cell Signaling Technology), anti-Akt antibody (1:1000) (#2920; Cell Signaling Technology), anti-VEGFR-2 antibody (1:1000) (#2479; Cell Signaling Technology), anti-VEGFR-1 (1:500) (#sc-316; Santa Cruz biotechnology), anti-β-actin antibody (1:5000) (#A2228; Sigma Aldrich), anti-Tie-2 antibody (1:500) (#MAB313; R&D systems) and anti-FGFR-1 antibody (1:1000) (#sc-121; Santa Cruz Biotechnology). Immunohistochemical staining were performed with anti-CD31 antibody (#Rb-10333-PO; Thermo Scientific). For immunofluorescence anti-CD31 (1:50) (# 553370; BD Pharmigen) and anti-VEGFR-2 antibody (1:50) (#2479; Cell Signaling Technology) were used. For flow cytometry, the following antibodies and dilutions were used: anti-CD31 (1:100) (#17-0319; eBioscience), anti-avb3 integrin (1:100) (MAB1976; Millipore), anti-VCAM-1(1:100) (#12-1069; eBioscience), anti-ICAM-1 (1:100) (#12-0549; eBioscience) and anti-MCP-1 (1:100) (#12-7099; eBioscience).

Faes S., Uldry E., Planche A., Santoro T., Pythoud C., Demartines N, & Dormond O. (2016). Acidic pH reduces VEGF-mediated endothelial cell responses by downregulation of VEGFR-2; relevance for anti-angiogenic therapies. Oncotarget, 7(52), 86026-86038.

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Histopathological and Immunohistochemical Analysis of Tumors

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Tumors were harvested and fixed with 4% formaldehyde at 4 °C for 24 h. Tissues were then embedded using paraffin and sectioned serially. The slides were dewaxed and stained with H&E for histopathological analysis. For IHC analysis, paraffin sections were deparaffinized, rehydrated, and subjected to antigen retrieval. Slides were then blocked with 10% normal goat serum and incubated with anti-Ki67 (1:500, Abcam, Cambridge, MA, USA), anti-CD31 antibody (1:100, Thermo Fisher Scientific), PTEN (1:100, Cell Signaling Technology, Beverly, MA, USA), or VEGF (1:100, Cell Signaling Technology) at 4 °C overnight, followed by an incubation with biotinylated secondary antibody and streptavidin-HRP. Immunoreactivity was visualized using AEC solution (Thermo Fisher Scientific).

Zhang H., Xu H.B., Kurban E, & Luo H.W. (2020). LncRNA SNHG14 promotes hepatocellular carcinoma progression via H3K27 acetylation activated PABPC1 by PTEN signaling. Cell Death & Disease, 11(8), 646.

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Isolation and Culture of Mouse Cardiac Endothelial Cells

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Mouse CEC were isolated with a modified protocol based on an online resource (http://vrd.bwh.harvard.edu/) and a publication [39 (link)]. Briefly, mouse hearts were dissected and minced into small pieces. After the digestion of the heart using Collagenase I (Worthington), cells were washed and incubated with Dynabeads conjugated with anti-CD31 antibody (Thermo Fisher). The beads with CEC were washed several times and cultured in a mouse endothelial culture medium (Cell Biologics). When confluent, cells were purified with Dynabeads conjugated with anti-Mouse CD102 (ICAM2) antibody. Each animal was used separate isolations, and cells less than passage 6 were used for the experiments.
For loss of function PGC-1α studies, we treated miR-21^+/+ or miR-21^−/− mouse CEC with either vehicle or a PGC-1α inhibitor SR-18292 (SR, MedChemExpress) at 20 μM for 72 h. SR-18292 induces PGC-1α acetylation [59 (link)]. Conversely, for the gain of function PGC-1α studies, we overexpressed PGC-1α in WT CEC using adenoviral Ad-PGC-1α and control Ad-GFP (generously provided by Dr. Pere Puigserver from Harvard Medical School) [65 (link)]. CEC were transduced with adenovirus at a multiplicity of infection (MOI) of 50 and treated with high glucose (25.5 mM) for 72 h.

Juguilon C., Wang Z., Wang Y., Enrick M., Jamaiyar A., Xu Y., Gadd J., Chen C.L., Pu A., Kolz C., Ohanyan V., Chen Y.R., Hardwick J., Zhang Y., Chilian W.M, & Yin L. (2022). Mechanism of the Switch from NO to H2O2 in Endothelium-Dependent Vasodilation in Diabetes. Basic research in cardiology, 117(1), 2.

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Histological and Immunofluorescence Analysis of Wound Healing

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After 12 days of treatment, wound tissues were excised, fixed in 10% formalin buffer at room temperature for 24 hours, and embedded in paraffin to prepare histological sections. The sections were stained with H&E and Masson’s trichrome. The bright-field microscopic images of the samples were recorded using an inverted fluorescence microscope (Olympus, IX83). Similarly, the wound tissues were collected on day 12 and were immunofluorescently stained with anti-CD31 antibody (Thermo Fisher Scientific, MA3105). Furthermore, the immunofluorescence staining of TNF-α and IL-6 were carried out by excising the wound tissues on day 9 and staining with primary antibodies anti–TNF-α antibody (Abcam, ab1793) and anti–IL-6 polyclonal antibody (Bioss, bs-0782R), respectively. All the steps of immunofluorescence staining were carried out according to the manufacturer’s protocol. The fluorescence intensities were measured using inverted fluorescence microscopy (Olympus, IX83).

Barman S.R., Chan S.W., Kao F.C., Ho H.Y., Khan I., Pal A., Huang C.C, & Lin Z.H. (2023). A self-powered multifunctional dressing for active infection prevention and accelerated wound healing. Science Advances, 9(4), eadc8758.

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Synthesis and Characterization of Chitosan-Coated Superparamagnetic Iron Oxide Nanoparticles

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ʟ-alanine and mPEG (Mn 2000) were purchased from Sigma-Aldrich (St Louis, MO, USA). Trifluoroacetic acid (TFA) and trifluoroacetic acid-d (TFA-d) were purchased from Alfa Aesar (Wall Hill, MA, USA). Ammonium hydroxide solution (28–30%), toluene, hexane, diethyl ether, and 95% ethanol were purchased from Echo Chemicals (Toufen, Miaoli, Taiwan). Dimethyl sulfoxide (DMSO) was purchased from J.T. Baker (Center Valley, PA, USA). Dulbecco’s modified Eagle’s medium (DMEM), fetal bovine serum (FBS), and antimycotic antibiotics were purchased from Gibco (Grand Island, NY, USA). Tetrahydrofuran (THF) was purchased from TEDIA (Fairfield, OH, USA); dichloromethane (DCM), N,N-dimethylformamide (DMF), and chloroform were obtained from AVANTOR (Center Valley, PA, USA) and dried over CaH2 before being used. Chitosan-coated superparamagnetic iron oxide (CSPIO) was prepared with a chemical co-precipitation method [29 (link)]. Prussian blue and anti-CD31 antibody were purchased from Thermo Fisher (Cheshire, UK). Anti-Ki67 antibody and anti-alpha smooth muscle actin (SMA) antibody were purchased from Abcam (Cambridge, UK).

Juang J.H., Chen C.L., Kao C.W., Chen C.Y., Shen C.R., Wang J.J., Tsai Z.T, & Chu I.M. (2023). The Image–Histology Correlation of Subcutaneous mPEG-poly(Ala) Hydrogel-Embedded MIN6 Cell Grafts in Nude Mice. Polymers, 15(12), 2584.

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Isolation of Mouse Cardiac Endothelial Cells

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Mouse cardiac endothelial cells (ECs) were isolated as previously described (20 (link)). Briefly, mouse hearts were dissected and minced into small pieces. After the digestion of the heart using Collagenase I (Worthington, Lakewood, NJ, United States), cells were washed and incubated with Dynabeads conjugated with anti-CD31 antibody (Thermo Fisher Scientific, Oakwood, OH, United States). The beads with endothelial cells were washed several times and cultured in a mouse endothelial culture medium (Cell Biologics, Chicago, IL, United States). When confluent, cells were purified with Dynabeads conjugated with anti-Mouse CD102 (ICAM2) antibody.

Winnicki A., Gadd J., Ohanyan V., Hernandez G., Wang Y., Enrick M., McKillen H., Kiedrowski M., Kundu D., Kegecik K., Penn M., Chilian W.M., Yin L, & Dong F. (2022). Role of endothelial CXCR4 in the development of aortic valve stenosis. Frontiers in Cardiovascular Medicine, 9, 971321.

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