Anti cd31

Manufactured by Thermo Fisher Scientific

Sourced in United States

Anti-CD31 is a laboratory product used for the detection and identification of CD31, a cell surface marker. It is commonly used in various cell biology and immunology research applications.

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Close spelling variants of this product

Anti-human CD31 (2 citations) Anti-CD31 mAb (2 citations) PE-conjugated anti-mouse PeCAM1 (CD31) antibody (2 citations) Anti-mouse CD31(PECAM-1)-PE (2 citations) APC-conjugated anti-CD31 (5 citations) Anti-CD31-FITC (8 citations) Anti-CD31 antibody-coated Dynabeads (2 citations) Mouse monoclonal anti-CD31 antibody (2 citations) Anti-CD31 antibody (15 citations) Anti-CD31-PE-Cyanine7 (WM-59, (2 citations)

47 protocols using anti cd31

Isolation and Characterization of PDLSCs

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Fourth-generation PDLSCs were collected and washed twice with PBS containing 1% FBS. After incubation in the dark with anti-CD45, anti-CD31, anti-CD29, anti-CD90 and anti-CD105 antibodies (Invitrogen, California, USA) at 4 °C for 30 minutes. The cells were centrifuged and washed three times, and the suspension was analyzed by sorted flow cytometry (Influx, BD, New Jersey, USA). The detection of DPSC surface antibodies was the same as above, and the markers included anti-CD73, anti-CD90, anti-CD45, anti-CD31 and Nestin (Invitrogen, California, USA).
Macrophage polarization markers were detected as follows. After washing with PBS, centrifuged cells with anti-F4/80 and anti-CD86 or anti-CD206 (Invitrogen, California, USA) were incubated in the dark at 4 °C for 30 minutes. The cells were then washed once, suspended in PBS containing 1% FBS and analyzed by flow cytometry. Flow cytometry data were analyzed by FlowJo software v10.2 (BD, New Jersey, USA).

Qiao X., Tang J., Dou L., Yang S., Sun Y., Mao H, & Yang D. (2023). Dental Pulp Stem Cell-Derived Exosomes Regulate Anti-Inflammatory and Osteogenesis in Periodontal Ligament Stem Cells and Promote the Repair of Experimental Periodontitis in Rats. International Journal of Nanomedicine, 18, 4683-4703.

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Tumor Cell Profiling by Flow Cytometry

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Cells were stained with a panel of antibodies in PBS containing 2% FBS, 1% penicillin-streptomycin, and 1 mM EDTA. Tumor cells from primary tumor samples were identified by gating out leukocytes, endothelial, and stromal cells expressing human lineage markers: CD2 (RPA-2.10), CD3 (UCHT1), CD18 (6.7), CD31 (WM59), CD45 (HI30), and CD64 (10.1). Tumor cells from xenografts passaged in mice were identified by gating out stromal cells expressing mouse lineage markers: H-2K^d (SF1–1.1), H-2K^b (AF6–88.5) and muCD45 (30–F11). Anti-CD31 was obtained from eBioscience, and all other antibodies were obtained from BD Pharmingen. All lineage antibodies were biotinylated or directly conjugated to PE/Cy5. Labeled cells were then washed and stained with streptavidin-PE/Cy5 (BioLegend). DAPI was obtained from Invitrogen.
After gating out leukocytes, endothelial, stromal, and non-viable cells, the tumor cells were profiled for expression of TLR2. Anti-TLR2 (T2.5) was obtained from eBioscience. Cells were analyzed on a BD LSRFortessa. Events collected were analyzed using FlowJo Version 9.6.4 software (Tree Star).

Farnebo L., Shahangian A., Lee Y., Shin J.H., Scheeren F.A, & Sunwoo J.B. (2015). Targeting Toll-like receptor 2 inhibits growth of head and neck squamous cell carcinoma. Oncotarget, 6(12), 9897-9907.

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Multimodal Immunolabeling of Diverse CNS Targets

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Catalogue numbers and concentrations of all antibodies are as follows. Anti-GFAP (130300, rat, 1:200), anti-occludin (711500, rabbit, 1:125) and anti-IgG (A11029, mouse, 1:100) were from Invitrogen. Anti–JAM-A (sc53623, mouse, 1:100) was from Santa Cruz Biotechnology. Fluoromyelin was from ThermoFisher (F34651, 1:300). Anti-fibrinogen (A0080, rabbit, 1:150) was from Dako. Anti-CD3 (16-0037-85, mouse, 1:100), anti-CD4 (14-9766-82, rat, 1:100), Anti-CD31 (550274, rat, 1:100), anti-CD45 (550539, rat, 1:100) were from eBioscience. anti-CD4 (ab183685, mouse, 1:50) was from Abcam. Anti-NeuN (MAB377, mouse, 1:100), anti-Olig2 (MABN50, mouse, 1:500) and anti-AQP4 (AB3594, rabbit, 1:200) were from Millipore. Anti-laminin (L9393, rabbit, 1:200) was from Sigma–Aldrich. Anti-Iba1 (109-19741, rabbit, 1:500) was from Wako.

Amatruda M., Chapouly C., Woo V., Safavi F., Zhang J., Dai D., Therattil A., Moon C., Villavicencio J., Gordon A., Parkos C, & Horng S. (2022). Astrocytic junctional adhesion molecule-A regulates T-cell entry past the glia limitans to promote central nervous system autoimmune attack. Brain Communications, 4(2), fcac044.

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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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Isolation of Lung Endothelial and Bone Marrow Monocytes

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Lungs were perfused with PBS and HBSS(+Collagenase 1 mg/ml; Sigma Aldrich, Switzerland) and minced before incubation with HBSS(+Collagenase) for 45 min and further processed for antibody incubation. Mouse lung cell suspension was incubated with anti-CD31(eBioscience, Austria, clone 390) antibody for 30 min in PBS (0.5% BSA) at 1 : 20 dilution in order to isolate lung ECs. To isolate monocytes from bone marrow cell suspension, the anti-CD115-biotin (eBioscience, clone AFS98) antibody was used at dilution of 1 : 50 in HBSS+2% FBS. Cell suspensions were incubated in primary antibody for 30 min at 4 °C followed by incubation with anti-rat IgG or streptavidin magnetic particles and sorted on LS columns according to the manufacturers recommendations (Miltenyi, Switzerland).

Hänggi K., Vasilikos L., Valls A.F., Yerbes R., Knop J., Spilgies L.M., Rieck K., Misra T., Bertin J., Gough P.J., Schmidt T., de Almodòvar C.R, & Wong W.W. (2017). RIPK1/RIPK3 promotes vascular permeability to allow tumor cell extravasation independent of its necroptotic function. Cell Death & Disease, 8(2), e2588-.

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Mouse Blood Cell Immunophenotyping

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Whole mouse blood was collected into microcentrifuge tubes containing 100μl HEPES medium (132mM NaCl, 6mM KCl, 1mM MgSO4, 20mM HEPES, and 5mM glucose) as previously described². Samples were diluted 2× in HEPES medium, and then centrifuged at RT for 15 min at 100 × g. Platelet rich plasma was incubated for 15 min at RT in 100μl of HEPES medium containing the following antibodies: Anti-CD45.2, anti-CD31, anti-CD9, anti-GARP and anti-Ter119 (eBioscience) prior to flow cytometry analysis. Data were acquired using BD Fortessa flow cytometers and analysed using FlowJo (TreeStar).

Nasrallah R., Imianowski C.J., Bossini-Castillo L., Grant F.M., Dogan M., Placek L., Kozhaya L., Kuo P., Sadiyah F., Whiteside S.K., Mumbach M.R., Glinos D., Vardaka P., Whyte C.E., Lozano T., Fujita T., Fujii H., Liston A., Andrews S., Cozzani A., Yang J., Mitra S., Lugli E., Chang H.Y., Unutmaz D., Trynka G, & Roychoudhuri R. (2020). A distal enhancer at risk locus 11q13.5 promotes suppression of colitis by Treg cells. Nature, 583(7816), 447-452.

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Immunostaining of Cutaneous Squamous Cell Carcinoma

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Tissue from freshly excised cSCC was obtained from patients during surgery at the Dermatology Department, University Hospital Southampton NHS Foundation Trust. The tissue samples (n=15 tumors) were snap frozen in liquid nitrogen, embedded in OCT medium, cryosectioned and immunostained as described previously (22 (link)). Primary antibodies included anti-CD3 (1:200, DAKO), anti-CD4 (1:50, Abcam), anti-CD8 (1:20, Invitrogen), anti-FOXP3 (1:20, eBioscience), anti-cytokeratin 16 (1:20, Thermo Scientific), anti-cytokeratin 17 (1:20, DAKO), anti CD31 (1:200, eBioscience), anti-CLA (1:200, Biolegend), anti-E-selectin (1:20, R & D Systems) and anti-OX40 (1:200, BD Biosciences). Fluorophore conjugated secondary antibodies comprised Alexa Fluor 488 goat anti-mouse IgG1a, Alexa Fluor 488 goat anti-rat IgM, Alexa Fluor 555 goat anti-rabbit IgG, Alexa Fluor 555 goat anti-rat IgG and Alexa Fluor 633 goat anti-mouse IgG2a (all from Invitrogen). Tissue sections were counterstained with DAPI (Sigma) before being mounted in Mowiol 4–88 (Harco) and imaged using a Zeiss Axioskop 2 fluorescence microscope or a Leica SP5 confocal microscope.

Lai C., August S., Albibas A., Behar R., Cho S.Y., Polak M.E., Theaker J., MacLeod A.S., French R.R., Glennie M.J., Al-Shamkhani A, & Healy E. (2016). OX40+ regulatory T cells in cutaneous squamous cell carcinoma suppress effector T cell responses and associate with metastatic potential. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(16), 4236-4248.

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Phenotypic Characterization of Cultured Cells

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CPs were stained for surface antigen expression using combinations of the following antibodies to confirm typical phenotype at P5: anti-CD31 (eBioscience, UK), anti-CD34 (eBioscience), anti-CD44 (eBioscience, UK), anti-CD45 (Miltenyi Biotec), anti-CD90 (BD Biosciences, UK), anti-CD105 (Life Technologies), and anti-CD146 (Miltenyi Biotec). After staining, fluorescence was analyzed using a FACS Canto II flow cytometer and FACS Diva software (both BD Biosciences, UK).

Avolio E., Rodriguez-Arabaolaza I., Spencer H.L., Riu F., Mangialardi G., Slater S.C., Rowlinson J., Alvino V.V., Idowu O.O., Soyombo S., Oikawa A., Swim M.M., Kong C.H., Cheng H., Jia H., Ghorbel M.T., Hancox J.C., Orchard C.H., Angelini G., Emanueli C., Caputo M, & Madeddu P. (2015). Expansion and Characterization of Neonatal Cardiac Pericytes Provides a Novel Cellular Option for Tissue Engineering in Congenital Heart Disease. Journal of the American Heart Association: Cardiovascular and Cerebrovascular Disease, 4(6), e002043.

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Multi-antibody Panel for Cell Sorting

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Monoclonal, murine-specific antibodies from BioLegend included: anti-CD11c (N418), anti-CD11b (M1/70), anti-Gr-1 (RB6-8C5), anti-CD19 (6D5), anti-CD4 (RM4–5), anti-CD8α (53-6.7), anti-CD25 (PC61), anti-NK1.1 (PK136), anti-Thy1.2 (53-2.1), anti-CD45.2 (104), anti-EpCAM (G8.8), and anti-CD45 (30-F11). Antibodies from eBioscience included: anti-CD11b (M1/70), anti-CD4 (RM4–5), anti-CD8α (53-6.7), anti-Thy1.2 (53-2.1), anti-F4/80 (BM8), anti-CD49b (DX5), anti-CD5 (53-7.3), anti-CD45 (30-F11), anti-IRF4 (3E4), anti-GATA3 (TWAJ), anti-B220 (RA3-6B2), and anti-CD31 (390). Antibodies from BD Biosciences included: anti-CD11c (HL3), anti-CD25 (PC61), anti-Thy1.2 (53-2.1), anti-Ly6G (1A8), anti-α4β7 (DATK32), and anti-Siglec-F (E50-2440). An Alexa Fluor 488-conjugated anti-Siglec-F antibody was generated using purified anti-Siglec-F with an Alexa Fluor 488 monoclonal antibody labeling kit (Life Technologies).
Exclusion of DAPI (4',6-diamidine-2'-phenylindole dihydrochloride; Roche) or LiveDead (Life Technologies) identified live cells, which were enumerated with flow cytometric CountBright Absolute beads (Life Technologies), according to the manufacturer’s instructions. Sample data were acquired with a LSRII flow cytometer (BD Biosciences) and analyzed using FlowJo software (Tree Star, Inc.). Cells were sorted using a MoFlo XDP (Beckman Coulter) instrument.

Mohapatra A., Van Dyken S.J., Schneider C., Nussbaum J.C., Liang H.E, & Locksley R.M. (2015). Group 2 innate lymphoid cells utilize the IRF4-IL-9 module to coordinate epithelial cell maintenance of lung homeostasis. Mucosal Immunology, 9(1), 275-286.

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Immunohistochemical Analysis of Solid Tumors

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Solid tumors were fixed with 10% phosphate buffered formalin, processed, and embedded in paraffin.
The sections were dewaxed and stained with H&E under a light microscope at 10× magnification. Immunohistochemistry (IHC) was performed according to the manufacturer’s instructions (LSAB kit; Dako, Carpinteria, CA, USA). Images were taken using a microscope. Cryosections of tumors were stained with anti-CD31 (eBioscience) and anti-phospho-Stat 3 (Santa Cruz) antibodies, respectively, followed by a biotinylated secondary antibody and streptavidin-FITC with DAPI counterstaining to detect tumor vasculature. The fluorescence images were taken by a microscope (Nikon Eclipse ci, Tokyo, Japan), and processed by using Image pro-plus 6.0 software.

Li W., Xue J, & Xu H. (2019). Combined administration of PTX and S-HM-3 in TPGS/Solutol micelle system for oncotarget therapy. International Journal of Nanomedicine, 14, 1011-1026.

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