PBMC staining and analysis were as described [3 (link)]. Unless otherwise specified, mAbs were from BD Pharmingen and included anti-CD3-Alexa Fluor 700 (UC-HT1), anti-CD4-APC (RPA-T4), anti-CD8-PacificBlue (RPA-T8), anti-CD11c-FITC (BU15, Thermo Scientific), anti-CD14-APC (RMO52, Beckman Coulter), anti-CD16-PacificBlue (3G8), anti-CD19-APC (HIB19) and anti-CD20-FITC (L27), anti-CD56-APC (N901, Beckman Coulter), and anti-NKG2D-PE (1D11; [2 (link)]). Anti-NKG2DL mAb were anti-MICA/B-PE (6D4), anti-ULBP1-PE (170818, R & D Systems), anti-ULBP2/5/6-PE (165903; R & D Systems), anti-ULBP3-PE (166510; R & D Systems), and anti-ULBP4-PE (1H1; [12 (link)]). In some cases, viable cell numbers were limiting resulting in partial data sets.
Anti ulbp2 5 6 pe
Manufactured by R&D Systems
Sourced in United States
Anti-ULBP2/5/6-PE is a fluorescently-labeled antibody that binds to the ULBP2, ULBP5, and ULBP6 proteins. It can be used for the detection and quantification of these proteins in various experimental applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti ulbp3 pe, by R&D Systems (3 mentions)
Anti cd56 apc, by Beckman Coulter (2 mentions)
Anti cd8 pacificblue, by Thermo Fisher Scientific (2 mentions)
Anti cd11c fitc bu15, by Thermo Fisher Scientific (2 mentions)
Anti cd14 apc rmo52, by Beckman Coulter (2 mentions)
Anti cd16 pacificblue 3, by Beckman Coulter (2 mentions)
Anti cd19 apc, by Beckman Coulter (2 mentions)
Anti mica b pe 6d4, by R&D Systems (2 mentions)
Anti ulbp1 pe, by R&D Systems (2 mentions)
Anti cd4 apc, by Thermo Fisher Scientific (2 mentions)
Anti cd20 fitc, by Beckman Coulter (2 mentions)
Anti cd3 alexa fluor 700 uc ht1, by Thermo Fisher Scientific (2 mentions)
Bicinchoninic acid assay, by Thermo Fisher Scientific (2 mentions)
7 aad staining, by BioLegend (1 mentions)
Cytomics fc500, by Beckman Coulter (1 mentions)
Apc conjugated annexin 5, by BD (1 mentions)
Anti p53 fl393, by Santa Cruz Biotechnology (1 mentions)
Anti cd45 hi30, by BD (1 mentions)
Anti mica, by R&D Systems (1 mentions)
Anti actin 1 19, by Santa Cruz Biotechnology (1 mentions)
4 protocols using anti ulbp2 5 6 pe
1
Multiparameter Immune Cell Analysis
2
Multiparameter Immune Cell Analysis
3
Evaluating MHC I and NKG2D Ligand Expression in Cancer Cell Lines
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HT-29, HCT 116, A2780 and A2780R were seeded at 5 × 104 cells/mL (1 mL in a 24-well plate) and rested overnight before treatment with Ara-C/DS (1 µg/mL of DS loaded with 50 nM of Ara-C), its components, or cGAMP (1 µg/mL) for 48 h. Cells were collected and stained with anti-HLA-ABC-FITC (MHC I, Biolegend, San Diego, CA, USA), anti-MICA/B-FITC (R&D Systems, Minneapolis, MN, USA) or anti-ULBP2/5/6-PE (R&D Systems, Minneapolis MN, USA) and analyzed using the Cytomics FC-500 (Beckman Coulter, Fullerton, CA, USA) where 104 events were measured and singlet live cells were gated. Dead cells were identified by 7-AAD staining (Biolegend, San Diego, CA, USA). Data was analyzed using the CXP Analysis Software.
4
Immune Profiling of NB Tumor Cells
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The following antibodies for flow cytometry were used: anti-CD107a-FITC (H4A3), anti-CD3-Alexa-700 (UCHT1), anti-CD56-PE-Cy7 (B159), and anti-CD45 (HI30), purchased from BD Biosciences; anti-ULBP1-PE (170818), anti-ULBP2/5/6-PE (165903), anti-ULBP3-PE (166510), anti-MICA (159227), anti-MICB (236511), anti-TRAIL/R2-APC (17908), anti-CD155/PVR-PE (300907), and anti-Nectin-2/CD112-APC (610603), purchased from R&D Systems; W6/32 which recognizes human fully assembled MHC class I heavy chains; and goat F(ab′)2 Fragment anti-mouse IgG FITC (IM1619, Dako) for flow cytometry. Apoptosis of tumor cells was evaluated with APC-conjugated AnnexinV (BD-Pharmingen) and propidium iodide (PI) (Sigma-Aldrich).
Flow cytometry was performed on FACSCantoII and analysed by FACSDiva Software (BD Biosciences).
ROS production was evaluated in drug-treated NB cell lines by using CellROX Deep Red Reagent (C10422, Invitrogen) and measured by flow cytometry.
Whole-cell extracts were quantified by a bicinchoninic acid assay (Thermo Fisher Scientific), resolved on 8–10% SDS-PAGE and electroblotted. Filters were probed with primary antibodies followed by goat anti-mouse and HRP-conjugated rabbit anti-goat IgG (Jackson). The following antibodies for Western blotting were used: anti-p53 (FL-393) and anti-actin (I-19), purchased by Santa Cruz Biotechnology.
Flow cytometry was performed on FACSCantoII and analysed by FACSDiva Software (BD Biosciences).
ROS production was evaluated in drug-treated NB cell lines by using CellROX Deep Red Reagent (C10422, Invitrogen) and measured by flow cytometry.
Whole-cell extracts were quantified by a bicinchoninic acid assay (Thermo Fisher Scientific), resolved on 8–10% SDS-PAGE and electroblotted. Filters were probed with primary antibodies followed by goat anti-mouse and HRP-conjugated rabbit anti-goat IgG (Jackson). The following antibodies for Western blotting were used: anti-p53 (FL-393) and anti-actin (I-19), purchased by Santa Cruz Biotechnology.
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