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Ulbp 1 170818

Manufactured by R&D Systems

ULBP-1 (170818) is a laboratory instrument designed for measuring biomolecular interactions. It utilizes label-free optical biosensing technology to detect and analyze the binding of analytes to immobilized ligands. The core function of this product is to provide researchers with a tool for real-time monitoring and quantification of molecular interactions.

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2 protocols using ulbp 1 170818

1

Phenotyping of Expanded Natural Killer Cells

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The phenotype of eNK cells was analyzed weekly using flow cytometry. The following mouse anti-human antibodies were used for staining: CD16 (3G8), CD56 (NCAM 16.2), NKG2D (1D11), DNAM-1 (DX11), CD3 (HIT3a) from BD PharMingen; CXCR1 (8F1), CXCR3 (CEW33D), CXCR4 (L276F12), CXCR6 (K041E5), CCR4 (L291H4), CCR5 (J418F1), CCR6 (G034E3), CCR9 (L053E8), CX3CR1 (2A9-1), Fas-L (NOK-1), IFNγ (B27) from BioLegend; CD62L (DREG-56), intracellular Perforin (dG9), Granzyme B (GB11), and TNF-α (MAb11) from eBioscience. For surface staining, NK cells were resuspended in FACS buffer (0.5% FBS and 2 mM EDTA in PBS), blocked with 10% mouse serum for 30 min, then incubated with the indicated antibodies for 30 min at 4°C using concentrations recommended by the manufacturers. For subsequent intracellular staining, NK cells were stained using the BD Cytofix/Cytoperm Kit following manufacturer protocol. Data was acquired using a BD FACS Accuri C6 and analyzed using FlowJo v10.2 software. Isotype IgG antibodies were used as negative controls. Human NK cells were defined as CD16+, CD56+ and CD3. Mouse anti-human HLA-ABC (W6/32), MICA/B (6D4), CD155 (2H7) and PD-L1 (MIH1) from eBioscience, ULBP-1 (170818) from R&D, and Fas (DX2) from BioLegend were used to determine MHC-I, NKG2D ligand and DNAM-1 ligand expression on TC106 with surface staining performed as above for NK cells.
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2

Comprehensive Immunophenotyping of Hematologic Malignancies

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Monoclonal antibodies (mAbs) were used to define blast populations as follows: AML: CD45 (BD, Hl30), CD34 (BD, 581/CD34), CD117 (Invitrogen,104D2), HLA-DR (BD, G46-6) and CD33 (Biolegend, WM53). T-ALL: CD45, CD5 (Biolegend, L17F12), CD7 (BD, M-T701), CD2 (BD, RPA-2.10), CD3 (BD, SK7) and CD8 (BD, RPA-T8). The following mAbs were used to define T cell subsets: CD3(BD, UCHT1), CD4 (BD, RPA-T4), CD8 (BD, RPA-T8)), CD45RO (BD, UCHL1), CD62L (BD, DREG-56), CD95 (Biolegend, DX2). RhNKG2D Fc chimera (R&D, 12990NK-050) and rhIgG1 Fc (R&D, 110-HG-100) were PE-conjugated per manufacturer instructions (Abcam) and titrated prior to use. For individual NKG2D-ligand detection the following mAbs were used: MICA (MBL, AMO1), MICB (MBL, BMO1) (both conjugated per manufacturer instructions, Abcam), ULBP-1 (170818), ULBP2/5/6 (165903), ULBP3 (166510), ULBP4 (709116) (all R&D). Cells were washed and stained in phosphate-buffered saline supplemented with 2% fetal calf serum (Gibco) at 4°C after blocking with FcR blocking reagent (Miltenyi). LIVE/DEAD® Fixable Aqua Dead Cell Stain (Invitrogen, L34966) was used for live/dead staining. When applicable, cells were fixed using Cytofix kits (BD). Flow cytometry was performed using 4-Laser M Fortessa Analyzers (BD). Flow cytometric analysis was performed using FlowJo V10 (Tree Star).
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