Cd90 pe 5e10

Manufactured by BD

CD90-PE (5E10) is a fluorescently-labeled monoclonal antibody that specifically binds to the CD90 (Thy-1) cell surface antigen. The CD90 antigen is expressed on a variety of cell types, including hematopoietic stem cells, mesenchymal stem cells, and certain subsets of T cells. The PE (phycoerythrin) fluorescent dye is conjugated to the antibody, allowing for the detection and analysis of CD90-positive cells using flow cytometry or other fluorescence-based techniques.

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CD90-PE (98 citations) CD90 (5E10) (6 citations) CD90 PE clone 5E10 (3 citations)

4 protocols using cd90 pe 5e10

Immunophenotyping of Mesenchymal Stem Cells

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AD-MSCs and freshly purified samples were harvested, washed twice in PBS, and then stained with conjugated antibodies against CD271-ALEXA FLUOR 647 (C40-1457, IgG1k; BD), CD44-FITC (G44-26, IgG2bk; BD), CD90-PE (5E10, IgG1k; BD), and CD45-FITC (5B1, IgG2a; Miltenyi), or with purified primary antibodies against CD29 (MAR4, IgG1k; BD), CD9 (M-L13, IgG1k; BD), and CD73 (AD2, IgG1k; BD). Where necessary, cells were then labeled with goat anti-mouse IgG-Alexa Fluor 488 secondary antibody (Invitrogen). Corresponding isotype-matched antibodies, CD3-Alexa Fluor 647 (UCHT1, IgG1k; BD), CD3-FITC (UCHT1, IgG1k; BD), CD3-PE (UCHT1, IgG1k; BD), CD8 (BW135/80, IgG2a; Miltenyi), and Purified CD3 (UCHT1, IgG1k; BD), were used as negative controls. Samples were acquired using an AccuriC6 (BD) flow cytometer. All data were analyzed by FlowJo software (Tree-Star).

Virzì F., Bianca P., Giammona A., Apuzzo T., Di Franco S., Mangiapane L.R., Colorito M.L., Catalano D., Scavo E., Nicotra A., Benfante A., Pistone G., Caputo V., Dieli F., Pirrello R, & Stassi G. (2017). Combined platelet-rich plasma and lipofilling treatment provides great improvement in facial skin-induced lesion regeneration for scleroderma patients. Stem Cell Research & Therapy, 8, 236.

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Immunophenotyping of FLT3-ITD AML Cells

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MNC from FLT3-ITD AML BM samples were stained for lineage markers using biotinylated antibodies: CD4 (RPA-T4; Biolegend), CD8a (RPA-T8; Biolegend), CD19 (HIB19; Biolegend), CD41 (MEM-06; Sigma), CD235alpha (HIR2; eBioscience), CD56 (B159; BD Pharmingen). Cells were then stained with the following fluorochrome-conjugated antibodies: CD34-FITC (581; BD Pharmingen), CD90-PE (5e10; BD Pharmingen), CD33-PC5.5 (D3HL60, Beckmann Coulter), CD45RA-APC Cy7 (H1100; BD Pharmingen), Streptavidin-eFluor 450 (eBioscience), CD38-APC (HB7; BD Pharmingen), CD45-APC-Cy7 (2D1; BD Pharmingen). PI was added as live/dead marker. Cell sorting was performed on a FACS Aria II (Becton Dickinson, Heidelberg, Germany). Sorting purity of >98% was routinely obtained.

Garz A.K., Wolf S., Grath S., Gaidzik V., Habringer S., Vick B., Rudelius M., Ziegenhain C., Herold S., Weickert M.T., Smets M., Peschel C., Oostendorp R.A., Bultmann S., Jeremias I., Thiede C., Döhner K., Keller U, & Götze K.S. (2017). Azacitidine combined with the selective FLT3 kinase inhibitor crenolanib disrupts stromal protection and inhibits expansion of residual leukemia-initiating cells in FLT3-ITD AML with concurrent epigenetic mutations. Oncotarget, 8(65), 108738-108759.

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Isolation and Characterization of Hematopoietic Stem and Progenitor Cells from Umbilical Cord Blood

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Anonymized human UCB samples were collected from healthy new-borns of both sexes at University Hospital Basel. Relevant ethical regulations were followed, according to the guidelines of the local Basel ethics committees (vote 13/2007V, S-112/2010, EKNZ2015/335). UCB cells were processed by density gradient centrifugation, and CD34+ cells were isolated using EasySep™ human cord blood CD34 positive selection kit II (cat# 17896, Stemcell Technologies, Vancouver, BC, Canada) and stored at -150 °C. Upon thawing, CD34+ cells were stained with the following antibodies: CD34 (APC conjugate, clone: 581, 1 : 100 dilution, BD Biosciences), CD38 (PEcy7, HB7, 1 : 200, BioLegend), CD45RA (FITC, HI100, 1 : 100, BioLegend), CD90 (PE, 5E10, 1 : 50, BD Biosciences) and CD49f (PEcy5, GoH3, 1 : 50, BD Biosciences) for 30 minutes at 4 °C. Cells were then washed and resuspended in PBS + 2% FCS with 1 μg ml -1 7-aminoactinomycin D (7-AAD, ThermoFisher Scientific). HSPCs, phenotypically defined as CD34+CD38-CD45RA -CD90+CD49f+, were sorted using a BD™ FACSAria III with 100 μm nozzle, purity mode and sorting purities ≥90%. Gates and thresholds were set according to fluorescence minus one (FMO) controls.

Dettinger P., Wang W., Ahmed N., Zhang Y., Loeffler D., Kull T., Etzrodt M., Lengerke C, & Schroeder T. (2020). An automated microfluidic system for efficient capture of rare cells and rapid flow-free stimulation. Lab on a chip, 20(22).

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Isolation and Characterization of Hematopoietic Stem Cells

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Cells were stained with LIVE/DEAD Fixable Blue Dead Cell Stain (Life Technologies) and then with anti-human CD34 PE-Cy7 (581, BioLegend; 1:100), CD38 Alexa Fluor 647 (AT1, Santa Cruz Biotechnologies; 1;50), CD45RA BV 421 (HI100, BD Biosciences; 1:25), and CD90 BV605 (5E10, BioLegend; 1:30) and analyzed by flow cytometry. For sorting of CD34⁺ or CD34⁺ CD38⁻ CD90⁺ cells, cord-blood-derived CD34⁺ HSPCs were stained directly after isolation from blood with anti-human CD34 FITC (8G12, BD Biosciences; 1:100), CD90 PE (5E10, BD Biosciences; 1:50), CD38 APC (HIT2, BD Bioscience; 1:50), and cells were sorted on a FACS Aria II (BD Bioscience).

Baik R., Cromer M.K., Glenn S.E., Vakulskas C.A., Chmielewski K.O., Dudek A.M., Feist W.N., Klermund J., Shipp S., Cathomen T., Dever D.P, & Porteus M.H. (2024). Transient inhibition of 53BP1 increases the frequency of targeted integration in human hematopoietic stem and progenitor cells. Nature Communications, 15, 111.

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