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Anti mouse cd31 clone mec 13

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Anti-mouse CD31 (clone MEC 13.3) is a monoclonal antibody used in flow cytometry and immunohistochemistry applications. CD31, also known as PECAM-1, is a cell surface glycoprotein expressed on endothelial cells, platelets, and various leukocyte subsets. This antibody can be used to detect and quantify CD31-positive cells.

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Lab products found in correlation

Ki 67 clone mib 1, by Agilent Technologies (2 mentions) αsma a2547, by Merck Group (1 mentions) Annexin 5 pacific blue, by Thermo Fisher Scientific (1 mentions) Facsdiva software version 6, by BD (1 mentions) Anti mouse cd8 clone 53 6, by Thermo Fisher Scientific (1 mentions) Anti foxp3 antibody clone fjk 16s, by Thermo Fisher Scientific (1 mentions) Anti mouse cd25 pc61, by Thermo Fisher Scientific (1 mentions) Anti mouse cd45 clone 30 f11, by BioLegend (1 mentions) Anti mouse pd l1 clone 10f 9g2, by BioLegend (1 mentions) Anti mouse cd4 clone rm4 5, by BD (1 mentions) Facscanto 2, by BD (1 mentions) Fixation permeabilization solution, by BD (1 mentions) Perm wash, by BD (1 mentions) Biotinylated secondary antibody, by Agilent Technologies (1 mentions)

5 protocols using anti mouse cd31 clone mec 13

1

Immunohistochemical Staining of Mouse Tissues

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Tumor sections were handled as described previously 9 (link). Sections were incubated with anti-mouse LYVE-1 polyclonal (Ab 14817, 1:200; Abcam, Cambridge, MA, USA) or monoclonal anti-α-smooth muscle actin Sigma (αSMA A2547, 1:1000; Sigma, France), and rat monoclonal anti-mouse CD31 (clone MEC 13.3, 1:1000; BD Pharmingen, Franklin Lakes, NJ, USA) antibodies.
Grépin R., Guyot M., Dumond A., Durivault J., Ambrosetti D., Roussel J.F., Dupré F., Quintens H, & Pagès G. (2020). The combination of bevacizumab/Avastin and erlotinib/Tarceva is relevant for the treatment of metastatic renal cell carcinoma: the role of a synonymous mutation of the EGFR receptor. Theranostics, 10(3), 1107-1121.
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2

Multiparametric Analysis of Mouse Blood and Tumor Cells

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Blood cells were collected from the eye vein of anaesthetised mice; splenocytes from disaggregated spleens were filtered through Falcon strainers. Mouse tumours were cut into small pieces, disaggregated with collagenase (0.5 mg ml−1) and filtered through strainers. Cells (106) were stained with live and death dye (Invitrogen, Carlsbad, CA, USA), and with the following specific antibodies: PerCp- Rat CD45 (30F11), APCRat CD11b (M1/70), PE Hamster CD3 (145-2C11), PE Rat Ly6C and Ly6C (RB6-8C5), FITC Rat Ly6C (AL-21), PE Rat Ly-6G (1A8), as well as relative control antibody Alexa Fluor 647 CD19 (1D3) (eBioscience, Waltham, MA, USA). PacificBlue Annexin V was purchased from Invitrogen, and streptavidin dye-conjugated (APC) and BD TruCOUNT tubes were obtained from BD Biosciences (San Jose, CA, USA). Flow cytometry was performed by FACS CantoTM instrument and FACS Diva software version 6.1.1 (BD Biosciences).
Immunohistochemistry of mouse tumours was performed with anti-mouse PE F4/80 (A3-1, Serotec (Oxford, UK), anti-mouse CD31 (clone MEC13.3; BD Biosciences). In each experiment, five to eight tumours per group were analysed; results are the mean of positive cells in four slices per tumour. To perform analysis we used ImageJ programme.
Belgiovine C., Bello E., Liguori M., Craparotta I., Mannarino L., Paracchini L., Beltrame L., Marchini S., Galmarini C.M., Mantovani A., Frapolli R., Allavena P, & D'Incalci M. (2017). Lurbinectedin reduces tumour-associated macrophages and the inflammatory tumour microenvironment in preclinical models. British Journal of Cancer, 117(5), 628-638.
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3

Quantification of Tumor Necrosis and Angiogenesis

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Sections of formol-fixed and paraffin-embedded tumors were stained with HES and necrotic area were quantified. Sections were incubated with anti-mouse CD31 (clone MEC 13.3, BD Pharmingen, diluted at 1:500) or Ki67 (clone MIB1, DAKO, Ready to use) antibodies.
Hagege A., Saada-Bouzid E., Ambrosetti D., Rastoin O., Boyer J., He X., Rousset J., Montemagno C., Doyen J., Pedeutour F., Parola J., Bourget I., Luciano F., Bozec A., Cao Y., Pagès G, & Dufies M. (2022). Targeting of c-MET and AXL by cabozantinib is a potential therapeutic strategy for patients with head and neck cell carcinoma. Cell Reports Medicine, 3(9), 100659.
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4

Mouse Tumor Single-Cell Immune Profiling

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Preparation of single-cell suspension of mouse tumor tissues by mechanical grinding. All single-cell suspensions were incubated with rat anti-mouse CD16/CD32 blocking antibody (4 μg/ml) for 15 min after thorough filtration and precipitation, stained with fluorescein-conjugated antibodies, multiple washed with PBS, and resuspended in 7-AAD (exclude non-viable cells). For anti-mouse CD4 (Clone RM4-5, BD Biosciences), anti-mouse CD8 (clone 53-6.7, eBioscience), anti-mouse CD45 (clone 30-F11, Biolegend), anti-mouse CD25 (PC61.5, eBioscience), anti-mouse PD-L1 (clone 10 F.9G2, BioLegend) and anti-mouse CD31 (clone MEC 13.3, BD Biosciences) staining, after incubation for 1 h, cells were washed with PBS for three times (1500 rpm, 5 min each), then detected by flow cytometry (BD FACSCanto II). For FoxP3 staining, after incubation, cells were washed and fixed with 1 ml of fixation & permeabilization solution (BD Biosciences) for 30–60 min, and washed twice with Perm Wash (BD Biosciences). Intracellular staining with anti-FoxP3 antibody (clone FJK-16s, eBioscience) was performed for 1 h. The next steps are the same as described above.
Liu S., Qin T., Liu Z., Wang J., Jia Y., Feng Y., Gao Y, & Li K. (2020). anlotinib alters tumor immune microenvironment by downregulating PD-L1 expression on vascular endothelial cells. Cell Death & Disease, 11(5), 309.
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5

Immunohistochemical Evaluation of Tumors

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Sections from blocks of formaldehyde-fixed and paraffin-embedded tumors were examined for immunostaining. Sections were incubated with monoclonal anti-mouse CD31 (clone MEC 13.3, BD Pharmingen, diluted at 1:500) or Ki67 (clone MIB1, DAKO, Ready to use) antibodies. Biotinylated secondary antibody (DAKO) was applied and binding was detected with the substrate diaminobenzidine against a hematoxylin counterstain.
Dufies M., Verbiest A., Cooley L.S., Ndiaye P.D., He X., Nottet N., Souleyreau W., Hagege A., Torrino S., Parola J., Giuliano S., Borchiellini D., Schiappa R., Mograbi B., Zucman-Rossi J., Bensalah K., Ravaud A., Auberger P., Bikfalvi A., Chamorey E., Rioux-Leclercq N., Mazure N.M., Beuselinck B., Cao Y., Bernhard J.C., Ambrosetti D, & Pagès G. (2021). Plk1, upregulated by HIF-2, mediates metastasis and drug resistance of clear cell renal cell carcinoma. Communications Biology, 4, 166.
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