Freshly prepared PBMCs were used. Subpopulations of T cells, B cells, natural killer (NK) cells and antigen-presenting cells (APC) were characterized by surface staining with fluorescence labelled anti-CD3, anti-CD4, anti-CD5, anti-CD8, anti-CD14, anti-CD16, anti-CD19, anti-CD25, anti-CD27, anti-CD38, anti-CD45RA, anti-CD45RO, anti-CD56 and anti-HLA-DR (BD Bioscience); anti-BDCA1, anti-BDCA2, anti-BDCA3, anti-BDCA4 and anti-slan (Miltenyi Biotec). Negative controls included directly labeled or unlabeled isotype-matched irrelevant antibodies (BD Biosciences). Freshly prepared CSF cells were used directly for FACS analysis. Fluorescence-labeled antibodies for surface staining were used as follows: anti-CD3, anti-CD4, anti-CD8, anti-CD14, anti-CD16, anti-CD19, anti-CD25, anti-CD27, anti-CD45RA, anti-CD45RO, anti-CD56 and anti-HLA-DR (BD Bioscience); anti-BDCA1 and anti-slan (Miltenyi Biotec). All cells were measured on a LSR-Fortessa (BD Biosciences) and evaluated by FACS-Diva Software (BD Bioscience).
Anti cd56
Manufactured by BD
Sourced in United States, Germany
Anti-CD56 is a laboratory reagent used for the identification and enumeration of Natural Killer (NK) cells in biological samples. It is a monoclonal antibody that binds to the CD56 antigen expressed on the surface of NK cells. The function of this product is to facilitate the detection and analysis of NK cells in research and clinical applications.
Automatically generated - may contain errors
Lab products found in correlation
Anti cd3, by BD (18 mentions)
Anti cd8, by BD (18 mentions)
Anti cd4, by BD (15 mentions)
Anti cd19, by BD (8 mentions)
Anti cd45ra, by BD (4 mentions)
Anti hla dr, by BD (4 mentions)
Anti cd45, by BD (4 mentions)
Anti cd3, by BioLegend (4 mentions)
Isotype matched irrelevant antibodies, by BD (3 mentions)
Anti cd45ro, by BD (3 mentions)
Facscalibur flow cytometer, by BD (3 mentions)
Anti nkg2d, by BD (3 mentions)
Anti cd107a, by BD (3 mentions)
Anti cd11c, by BD (3 mentions)
Anti cd69, by BD (3 mentions)
Anti cd94, by BD (3 mentions)
Facsdiva software, by BD (2 mentions)
Lsrfortessa, by BD (2 mentions)
Anti bdca2, by Miltenyi Biotec (2 mentions)
Anti slan, by Miltenyi Biotec (2 mentions)
39 protocols using anti cd56
1
Comprehensive Immune Cell Profiling
2
GD2 Expression Analysis in Human NB Cells
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Purified mouse anti‐human GD2 mAb 14.G2a and purified goat anti‐mouse IgG Abs (BD Pharmingen, Franklin Lakes, NJ, USA) were used to detect the GD2 expression by human NB cell lines. The other mAbs used were: FITC‐labeled anti‐CD3, anti‐CD14 (BD Pharmingen), anti‐TCR Vα24, PE‐labeled anti‐TCR Vβ11, (Beckman Coulter, Brea, CA, USA), anti‐CD1d, APC‐labeled anti‐CD3, anti‐CD56, PE‐Cy7‐labeled anti‐CD8, anti‐CD56, PB‐labeled anti‐CD4, anti‐CD3 (BD Pharmingen), anti‐CD16 (BioLegend, San Diego, CA, USA), anti‐human TNF‐α, IFN‐γ, GM‐CSF blocking Abs (BioLegend) and anti‐human IL‐2 blocking Abs (R&D Systems, Minneapolis, MN, USA). The surface phenotypes of PBMC (peripheral blood mononuclear cells) and cultured cells were determined by a FACSCantoII instrument (BD Biosciences, Franklin Lakes, NJ, USA) and were analyzed using the FlowJo software program (Tree Star, Ashland, OR, USA).
3
Multiparametric Flow Cytometry Analysis
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Primary antibodies: anti-MET (Human HGFR/c-MET APC-conjugated Antibody, clone 95,106, R&D System); anti-IgG1/CH2CH3 regions (Alexa Fluor® 647 AffiniPure F (ab’)2 Fragment Goat Anti-Human IgG (H + L) antibody, Jackson Immuno Research); anti-CD4 (APC Mouse Anti-Human CD4 clone M-T466, Miltenyi); anti-CD3 (PE Mouse Anti-Human CD3, Clone HIT3a); anti-CD8 (APC Mouse Anti-Human CD8, Clone RPA-T8); anti-CD56 (PE Mouse Anti-Human CD56, Clone MY31) all from BD Biosciences. Isotype control antibodies: APC, FITC, or PE mouse IgG1 κ Isotype Control, Clone MOPC-21 (BD Biosciences). Cells were counterstained with DAPI and analyzed by Cyan ADP flow cytometer (Beckman Coulter S.r.l.). Data were elaborated using Summit 4.3 software (Dako). For plots in which the isotype control is not shown, the Mean Fluorescence Intensity (MFI) derived from the Isotype control was set within the first logarithm (0 < MFI < 10). Cells were considered positive for the analyzed marker if the signal was higher than 10 (MFI > 10).
4
Lymphocyte Subset Analysis by Flow Cytometry
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CD3 represents the total T cell number (thymus dependent lymphocyte) and CD19 is a cell surface receptor complex component that regulates B lymphocyte responses47 (link). Cells from whole blood samples were subjected to flow cytometry analysis to determine the proportions of lymphocyte subsets in peripheral venous blood within 2 h of collection, based on binding of the following monoclonal antibodies (mAbs): Anti-CD4, anti-CD8, anti-CD3, anti-CD56 and anti-CD19 (BD Biosciences, San Jose, CA, USA), The stained cells were sorted using a FACS Aria flow cytometer (BD Biosciences) using appropriate isotype controls for gating, and the data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA).
5
Flow Cytometric Analysis of T Cell Activation and Apoptosis
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To control the purity of cell selection, the following antibodies (BD Pharmingen™, Franklin Lakes, NJ, USA) were used: anti-CD3, anti-CD4, anti-CD8, anti-CD14, anti-CD19 and anti-CD56. Antibodies used for TNF receptor expression (anti-TNFR1 and anti-TNFR2) were purchased from R&D Systems (R&D Systems). To bring out the T cell activation, the following antibodies (Beckman Coulter, Brea, CA, USA) were used: anti-CD3, anti-CD25, anti-HLA-DR. To bring out the early T cell activation, the marker CD69 was studied after TNFα stimulation by flow cytometry (BD Pharmingen™). To detect apoptosis in T cells after TNFα stimulation, a fluorescein isothiocyanate (FITC) annexin-V Apoptosis Detection Kit (Becton-Dickinson, Franklin Lakes, NJ, USA) was used. Appropriate isotype controls were used in all cases. Acquisition of samples was performed on a FACSCanto flow cytometer (Becton-Dickinson), and the data were analyzed with FlowJo software.
6
Immunophenotyping and Cytokine Analysis
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Freeze-dried BCG (Chengdu Institute of Biological Products, Chengdu, China) was reconsitituted in a 0.9% sodium chloride solution to a concentration of 1 mg/mL prior to use. Sonicatd M.tb protein antigen (TB-Ag) was generously provided by Prof. Baiqing Li (Department of Immunology, Bangbu Medical College, Anhui Key Laboratory of Infection and Immunity, Bangbu, China). The following mAbs were used for phenotypic and intracellular cytokine analysis: phycoerythrin (PE)-labeled anti-CD3, anti-CD14, anti-CD56, anti-CD45RA, anti-IL-17, fluorescein isothiocyanate (FITC)-labeled anti-CD3, anti-CD45RA, anti-CD45RO, anti-granzyme B, Phycoerythrin-Cy7 (PE-cy7)-labeled anti-CD69, anti-CD56, allophycocyanin (APC)-labeled anti-CD3, anti-IFN-γ anti-IL-22 were obtained from BD Pharmingen (San Jose, CA, USA). FITC-labeled anti-CD16 was purchased from Biolegend (CA, USA). PE-labeled anti-NKG2D were obtained from R&D Systems (MN, USA), respectively.
7
Flow Cytometric Phenotyping of Immune Cells
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The following conjugated antibodies were used for flow cytometric phenotyping and analysis: anti-CD34, anti-CD43, anti-KDR, anti-CD45, anti-CD7, anti-CD5, anti-CD4, anti-CD8, anti-CD3, antiTCRαβ, anti-CD56, anti-CD15, anti-CD14, and anti-CD235a purchased from BD Biosciences. All antibodies were used in a 1:30 dilution. Dead cells were excluded from analysis in all experiments by staining with DAPI. Flow cytometry analysis was conducted on an LSRII cytometer (BD Biosciences, Paris, France), a FACS CANTO (BD Biosciences, Paris, France), or a FACS CELESTA (BD Biosciences, Paris, France) and analyzed using FlowJo software (BD Biosciences, Ashland, OR, USA).
8
Multiparameter Immunophenotyping of PBMCs
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9
NK Cell Degranulation Assay for AML
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Tested items and PBMCs from patients with AML were added to each well of round-bottomed 96-well plates. After overnight coincubation with the NKCE molecules or antibodies, antihuman CD107a and CD107b antibodies (Miltenyi, 130-111-621 and 130-118-818) were added for 4 h. Cells were then washed and stained with the following mixture: viability markers, anti-CD45 (Miltenyi, 130-110-771), anti-CD33 (BD Biosciences, 564588), anti-CD56 (BD Biosciences, 557747) and anti-CD3 (BD Biosciences, 740187) antibodies. Cells were then washed, fixed and analyzed by flow cytometry. The data obtained were analyzed with Flowjo Software to assess NK cell degranulation by monitoring the expression of CD107a/b on NK cells identified as living CD45+CD33−CD56+CD3− cells.
10
Immune Receptor Repertoire Analysis
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