PEC, PEC/hCD177, and PEC/hCD31 were plated in flat bottom gelatin-coated 24-well dishes at a concentration of 6 × 104/well. After culturing for 24 h, neutrophils from peripheral blood were applied to each well at a concentration of 3 × 105/well in the medium of 200 nmol/L phorbol 12-myristate 13-acetate (PMA). After a 2-h incubation, cells were obtained and were fluorescently labeled with allophycocyanin-conjugated anti-CD11b antibody (Bio Legend), Annexin V (Bio Legend), and 7-AAD (MBL Life science, Nagoya, Japan), CD11b-negative cells are gated as the target cells. The % cytotoxicity was calculated as (% Annexin V+ 7-AAD+ cells in CD11b+ cells) + (% Annexin V+ 7-AAD– cells in CD11b+ cells) + (% Annexin V– 7-AAD+ cells in CD11b+ cells). For the blocking assay, neutrophils were treated with 200 μg/mL of antihuman CD31 (clone: WM59, Bio Legend) or isotype mouse IgG1 (Santa Cruz, Dallas, TX) for 2 h at room temperature before adding to each well.
Antihuman cd31
Manufactured by BioLegend
Sourced in United States
The Antihuman CD31 is a lab equipment product that is used for the detection and identification of CD31, also known as PECAM-1, a glycoprotein expressed on the surface of platelets, monocytes, neutrophils, and endothelial cells.
Automatically generated - may contain errors
Lab products found in correlation
Annexin 5, by BioLegend (1 mentions)
Anti tnf α, by Thermo Fisher Scientific (1 mentions)
Ccl22, by R&D Systems (1 mentions)
Recombinant murine ccl19, by R&D Systems (1 mentions)
Anti laminin, by Abcam (1 mentions)
Anti human vegfr3, by BioLegend (1 mentions)
Alexfluor 555 phalloidin, by Thermo Fisher Scientific (1 mentions)
Anti collagen, by Abcam (1 mentions)
Cxcl12, by R&D Systems (1 mentions)
Cxcl10, by R&D Systems (1 mentions)
Sphingosine 1 phosphate (s1p), by Avanti Polar Lipids (1 mentions)
Tnf α, by R&D Systems (1 mentions)
Anti β catenin, by Abcam (1 mentions)
Interleukin 6 (il 6), by R&D Systems (1 mentions)
Ifn γ, by R&D Systems (1 mentions)
Anti ve cadherin, by BD (1 mentions)
3 protocols using antihuman cd31
1
Neutrophil Cytotoxicity Assay on PEC Cells
2
Lymphatic Endothelial Cell Markers
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S1P was from Avanti Polar Lipids (Alabaster, AL). Recombinant murine CCL19, CCL5, CCL22, CCL2, CXCL12, CXCL10, IL-6, TNFα and IFNγ were from R&D Systems (Minneapolis, MN). Anti-VCAM-1 (clone 429), anti-ICAM-1 (clone YN1/1), anti-CD4 (GK1.5), anti-CD25 (clone PC61.5), anti-CD44 (clone IM7) and anti-TNFα were from eBioscience (San Diego, CA). Anti-human CD31, anti-human LYVE1, anti-human VEGFR3, anti-human GP38, anti-human ICAM-1 and anti-human VCAM-1 antibodies were from Biolegend (San Diego, CA). Anti-VE-cadherin was from BD PharMingen (San Diego, CA). AlexFluor-555 phalloidin was from Life Technologies (Grand Island, NY). Anti-moesin was from Cellsignaling (Danvers, MA). Anti-β-catenin, anti-collagen and anti-laminin were from Abcam (Cambridge, UK). Anti-prox1 antibody was from Bioss (Woburn, MA). Recombinant human laminin 511 was from Biolamina AB (Sundbyberg, Sweden). EIA grade gelatin was from Bio-Rad (Berkely, CA).
3
Immunohistochemical Analysis of Implant Vascularization
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Mice were euthanized, and both the implant and controls ears were cut off and placed in PBS. Using forceps, the skin on the ventral side of the ear was retracted to expose the human tissue implant. The ears were fixed in 4% paraformaldehyde for 20 minutes at 4 °C and incubated overnight in 0.05% Triton X-100 in PBS containing anti-human CD31 (Biolegend, San Diego, CA, USA) antibody conjugated to either PE or AF647 in a 1:200 dilution. The tissue was washed in PBS and mounted on a glass slide with a glass coverslip for imaging.
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