Bone marrow aspirates from patients with MGUS and MM were centrifuged on Ficoll- Hypaque (Pharmacia Biotech, Uppsala, Sweden) gradient, and the separated mononuclear cells were left to adhere in complete medium (RPMI-1640 medium supplemented with 10% FBS). To isolate ECs, BM adherent stromal cells were harvested and incubated with magnetic microbeads coated with anti-CD31 antibody (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s instructions. Factor VIII-related antigen, CD31, Vascular Endothelial Growth Factor Receptor 2 (VEGFR2), and Tie2 expression of freshly isolated MGUS-ECs and MM-ECs was verified by flow cytometry FACS Canto II (Becton Dickinson-BD, San Jose, CA, USA).
Anti cd31 antibody
Manufactured by Miltenyi Biotec
Sourced in Germany
The Anti-CD31 antibody is a laboratory tool used for the identification and analysis of CD31-positive cells. CD31, also known as PECAM-1, is a cell surface glycoprotein expressed on endothelial cells, platelets, and certain immune cells. The Anti-CD31 antibody can be used in various cell-based assays and applications, such as flow cytometry, immunohistochemistry, and Western blotting, to detect and quantify CD31-expressing cells.
Automatically generated - may contain errors
Lab products found in correlation
Ficoll hypaque, by GE Healthcare (2 mentions)
Facscanto 2, by BD (2 mentions)
Kaluza acquisition software, by Beckman Coulter (1 mentions)
Anti sca 1, by Miltenyi Biotec (1 mentions)
Anti cd117, by Miltenyi Biotec (1 mentions)
Gallios flow cytometer, by Beckman Coulter (1 mentions)
Magnetic column, by Miltenyi Biotec (1 mentions)
Neural tissue dissociation kit p, by Miltenyi Biotec (1 mentions)
4 protocols using anti cd31 antibody
1
Isolation and Characterization of Endothelial Cells from MGUS and MM
2
Phenotypic Analysis of Cultured hCSCs
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Cultivated hCSCs were harvested by centrifugation after treatment with trypsin and subsequently stained with PE-coupled anti-CD105, anti-CD117, anti-Sca1 or anti-CD31 antibody (Miltenyi Biotec, Bergisch Gladbach, Germany) according to manufacturer’s guidelines. For isotype controls, hCSCs were stained with PE-coupled IgG1 control antibody or APC-coupled IgG1 control antibody. Analysis was done using Gallios Flow Cytometer (Beckmann Coulter Inc., Brea, CA, USA), while Kaluza Acquisition Software (Beckmann Coulter Inc.) was used for subsequent data acquisition and statistical analysis.
3
Isolation and Characterization of Myeloma Endothelial Cells
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BM mononuclear cells from MM and MGUS patients were obtained by centrifugation on Ficoll-Hypaque (Pharmacia Biotech, Uppsala, Sweden) gradient of heparinized BM aspirates. Cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic/antimycotic. To isolate ECs, BM adherent stromal cells were harvested and incubated with magnetic microbeads coated with anti-CD31 antibody (Miltenyi Biotec, Bergisch Gladbach, Germany), according to manufacturer's instructions. Factor VIIIerelated antigen, CD31, vascular endothelial growth factor receptor 2, and tyrosine kinase with immunoglobulin-like and EGF-like domains 2 (Tie2) expression levels on freshly isolated endothelial cells from MGUS (MGECs) and endothelial cells from MM (MMECs) patients were analyzed by flow cytometry, FACS Canto II (Becton Dickinson, San Jose, CA).
4
Isolating CD31+ Endothelial Cells from Mouse Brain
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P5 mice were knocked down using ice and decapitated. Brains were removed and placed in ice cold PBS. Enzymes from the Neural Tissue Dissociation Kit (P) (MACS Miltenyi Biotec) were added to the brains and were then digested using the gentleMACS Octo Dissociator with Heaters following the manufacturer’s protocol. The dissociated cell suspension was then incubated with beads conjugated with anti-CD31 antibody (MACS Miltenyi Biotec) and applied to a magnetic column (MACS Miltenyi Biotec). CD31+ cells were eluted from the column for RNA isolation.
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