Cells were stained with anti-human CD3-APC, anti-human CD4-PE, anti-human CD8-FITC, anti-human CD56-PE, anti-human CD279-APC, and anti-human CD137-PE (BD Biosciences, USA) for 15 mins at 4°C. They were then washed with and re-suspended in PBS. Events were acquired using a flow cytometer (Calibur, BD Bioscience) and analyzed by Cell Quest software (BD Biosciences).
Anti human cd56 pe
Manufactured by BD
Sourced in United States
The Anti-human CD56-PE is a monoclonal antibody conjugated with the fluorescent dye phycoerythrin (PE). It is designed for the detection and identification of CD56-positive cells, which are typically found on natural killer (NK) cells and a subset of T cells. The antibody can be used in flow cytometry applications to analyze and quantify CD56-expressing cell populations.
Automatically generated - may contain errors
Lab products found in correlation
Ficoll hypaque, by GE Healthcare (3 mentions)
Anti cd3 percp, by BioLegend (3 mentions)
Golgistop, by BD (2 mentions)
Anti cd107a alexa fluor 647 antibody, by Thermo Fisher Scientific (2 mentions)
Anti human cd158b fitc, by BD (2 mentions)
Anti human cd3 apc, by BD (2 mentions)
Anti human cd3 fitc, by BD (2 mentions)
Facscalibur flow cytometer, by BD (2 mentions)
Anti human cd4 pe, by BD (1 mentions)
Anti human cd8 fitc, by BD (1 mentions)
Cellquest software, by BD (1 mentions)
Calibur, by BD (1 mentions)
Recombinant human il 15, by R&D Systems (1 mentions)
Anti cd107a efluor 660 antibody, by Thermo Fisher Scientific (1 mentions)
Versacomp antibody capture bead kit, by Beckman Coulter (1 mentions)
Pe cy7, by BD (1 mentions)
Anti human cd2 pe, by BD (1 mentions)
Anti human cd15 pe, by BD (1 mentions)
Anti human cd19 pe, by BD (1 mentions)
Lsrfortessa cell analyzer, by BD (1 mentions)
11 protocols using anti human cd56 pe
1
Multiparametric Flow Cytometry Analysis
2
PBMC Cytotoxicity Assay with IL-15 Stimulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human PBMC were isolated from the blood of 8 healthy donors using Hypaque-Ficoll (GE Healthcare, Amersham, UK) density centrifugation, with informed consent and full ethical approval (NRES reference: 06/Q1701/120). 3 × 105 PBMCs were stimulated overnight with 1 ng/mL recombinant human IL-15 (R&D Systems). Peptide pulsed 721.221C*0304-ICP47 targets were prepared as for the stabilization assays. Target cells were resuspended with PBMCs at an effector-to-target (E:T) ratio of 5:1 in fresh R10 medium containing peptide and anti-CD107a-efluor-660 antibody (eBioscience, Hatfield, UK). Cells were incubated for 1 h at 26°C, then 6 μg/mL Golgi- Stop™ (BD Biosciences) was added, and incubated for a further 4 h at 26°C. Cells were washed, blocked with blocking buffer for 30 min and then stained with the following antibodies: anti-CD3-PerCP (Biolegend, San Diego, USA), anti-human CD56-PE, and anti-human KIR2DL2/L3/S2, CD158b-FITC (both BD Biosciences). Cells were fixed in 1% PFA and analyzed by flow cytometry. Individual assays for each donor were performed once in duplicate and the mean value used for subsequent analysis.
3
Immunophenotyping of Blood Cell Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The antibodies used for differentiation in the described method were anti-human CD14 Pacific Blue® (Clone: M5E2), anti-human CD16 phycoerythrin-cyanine 7 (PE-Cy 7, Clone: 3G8), and anti-human HLA-DR allophycocyanin and fluorescein isothiocyanate (APC, FITC, Clone: TU36), and negative lineage markers anti-human CD2 PE (Clone: RPA-2.10), anti-human CD15 PE (Clone: VIMC6), anti-human CD19 PE (Clone: HIB19), anti-human CD56 PE (Clone: MY31), and anti-human NKp46 PE (Clone: BAB281) from BD Biosciences, Franklin Lakes, NJ, USA. For fluorescence compensation, the VersaComp antibody capture bead kit from Beckman Coulter, Inc., Brea, CA, USA was used. All flow cytometry-based assays were analyzed on BD LSR Fortessa™ cell analyzer.
4
Longitudinal Immune Cell Profiling in aHSCT
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We collected whole blood samples from patients before aHSCT (pre) with varying timepoints before mobilization chemotherapy and on the following timepoints after HSCT (in months): 1, 3, 6, 9, 12, 18, 24, 30, 36, 42, 48, 54, 60. Samples collected after 5 years after aHSCT were summarized (post).
Lymphocyte immunophenotyping of samples was performed with BD FACSCalibur™ using conjugated antibodies from BD Biosciences™: anti-human CD3 FITC (SK7), anti-human CD45 PerCP (2D1), anti-human CD8 PE (SK1), anti-human CD4 APC (SK3), anti-human CD19 APC (2SJ25C1), anti-human CD16 PE (B73.1), anti-human CD56 PE (NCAM16.2) and FITC Mouse IgG1, κ Isotype Control.
Lymphocyte immunophenotyping of samples was performed with BD FACSCalibur™ using conjugated antibodies from BD Biosciences™: anti-human CD3 FITC (SK7), anti-human CD45 PerCP (2D1), anti-human CD8 PE (SK1), anti-human CD4 APC (SK3), anti-human CD19 APC (2SJ25C1), anti-human CD16 PE (B73.1), anti-human CD56 PE (NCAM16.2) and FITC Mouse IgG1, κ Isotype Control.
5
Multiparametric Flow Cytometry of Immune Cell Populations
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peripheral blood mononuclear cells (PBMCs) were isolated from EDTA blood using a Ficoll gradient and frozen (− 80 °C) until further analysis. For flow cytometry, the following antibodies were used: anti-human CD3-APC (1/100) clone UCHT1, anti-human CD4-FITC (1/100) clone SK3, anti-human CD8-PerCP (1/100) clone SK1, anti-human CD19-FITC (1/100) clone HIB19, anti-human CD27-PE (1/50) clone L128, anti-human CD38-APC (1/50) clone HIT2, anti-human CD56-PE (1/50) clone B159, anti-human CD197(CCR7)-PE (1/50) clone 3D12 (all BD Biosciences), anti-human CD45RA-APC (1/50) clone HI100 (BioLegend). As a reference, analyses were additionally performed in 10 healthy controls. Gating strategy for CD4/CD8-T cell subsets, NK cells and B cell-subsets is displayed in Supplementary Fig. S1 .
6
Phenotypic Characterization of Cytokine-Induced Killer Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The expression of CIK CD markers on the CIK cells was evaluated by flow cytometry on the 15th day. For this purpose, 2 × 105 CIK cells were washed once with PBS, and then were suspended in a tube containing PBS containing 3% bovine serum albumin (Sigma, A7284-10ML). Immediately, each tube was incubated at 4oC with anti-human CD3-PerCP (Miltenyi Biotec, 130-094-965) or anti-human CD56-PE (BD, BD 561,903), and appropriate isotype controls [Mouse IgG2a isotype PerCP (Miltenyi Biotec, 130-099-190) and mouse IgG1 isotype PE (BD, BD 556,650)]. Next, the mixture was incubated at 4 °C and washed twice with PBS. In addition, the cell pellet was washed twice and then were suspended in PBS. The cell phenotype was then analyzed using BD FACSCalibur flow cytometer (BD Biosciences, USA), and the resultant flow cytometry data were analyzed using. Flowjo 8 software (FlowJo LLC, USA) was used to analyze flow cytometry data.
7
Quantifying NK cell activation by IL-15
8
Cytotoxicity Assay of Human PBMC Stimulated with IL-15
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Human PBMC were isolated from the blood of 14 healthy donors using Hypaque‐Ficoll (GE Healthcare, Amersham, UK) density centrifugation, with informed consent and full ethical approval (REC ref 06/Q1701/120). A total of 3 × 105 PBMCs were stimulated overnight with 1 ng/mL recombinant human IL‐15. Peptide pulsed 721.174 targets were prepared as for the stabilization assays. Target cells were resuspended with PBMCs at an effector‐to‐target (E:T) ratio of 5:1 in fresh R10 medium containing peptide and anti‐CD107a‐Alexa‐fluor‐647 antibody (eBioscience, Hatfield, UK). Cells were incubated for 1 h at 26°C, then 6 μg/mL Golgi‐StopTM (BD Biosciences) was added, and incubated for a further 3 h at 26°C. Cells were washed in wash buffer (1% BSA/0.1% NaN3) and blocked with blocking buffer (10% human serum in PBS) for 30 min and then stained with the following antibodies: anti‐CD3‐PerCP (Biolegend, San Diego, USA), anti‐human CD56‐PE, and anti‐human CD158b‐FITC (both BD Biosciences). Cells were fixed in 1% PFA and analyzed by flow cytometry. Individual assays for each donor were performed once in duplicate and the mean value used for subsequent analysis.
9
CAR T Cell Phenotypic Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell proliferation was determined by counting the cells using automated cell counter R1 (Olympus, Tokyo, Japan). The expression of CAR on the cell surface was determined using phycoerythrin (PE) anti‐human GM‐CSF and PE anti‐human CD33 (both from Miltenyi Biotec); allophycocyanin (APC) anti‐human CD3, peridinin–chlorophyll–protein (PerCP) anti‐human CD4, PerCP anti‐human CD8, fluorescein isothiocyanate anti‐human CD45RA, PE anti‐human CD56 and PE anti‐human CD62L (all from BD Pharmingen); PE anti‐human CD116 (Beckman Coulter, Pasadena, CA); and APC anti‐human CD197 [C‐C chemokine receptor type 7 (CCR7)] and APC anti‐human programmed cell death 1 (PD‐1; both from BioLegend, San Diego, CA) to characterise the phenotype of CAR T cells and examine the cell lines for coculture experiments. All flow cytometry data were acquired using BD fluorescent‐activated cell‐sorting Accuri C6 Plus and analysed using FlowJo v10.5 software (both from BD Biosciences, Franklin Lakes, NJ).
10
Flow Cytometry Analysis of NK Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The previous study showed that the NK cells were analyzed with the help of a marker on flow cytometry [12 (link)]. In this study, 200 μL heparinized whole blood samples were surface-stained with 5 μL FITC anti-human CD3 (BD Biosciences, 561806, USA), APC anti-human CD16 (Zen Biosciences, 790365, CN), PE anti-human CD56 (BD Biosciences, 561903, USA), and isotype control for 30 min at room temperature, respectively. The red cells were then lysed with red cell lyse buffer (BD, USA) for 30 min at room temperature and washed twice with 500 μL phosphate buffer solution (PBS) via centrifugation. The resultant white cell pellets were resuspended and analyzed with a FACSCalibur flow cytometer (Becton Dickinson, San Jose, CA, USA). Lymphocytes were gated, and NK cell population was analyzed for the immunophenotypic features as CD3−CD16+CD56+ NK cells.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!