For staining of peripheral blood mononuclear cells with purified antibodies, 100,000 cells were incubated with 0.2μg of antibody diluted in PBS/0.5% BSA/0.02% NaN3 (FACS medium) for 1h on ice as previously described [42 (link), 43 (link)]. Cells were centrifuged 400g/5min and supernatant was removed. The cells were incubated 30mins on ice with secondary antibodies, washed with FACS medium and analyzed with FACS calibur instrument (BD, Franklin Lakes, NJ, USA) and FlowJo or CellQuest software. The following antibodies were used: PE-anti-CD56 (DAKO, Glostrup, Denmark, clone MOC-1), FITC-anti CD3 (DAKO, clone UCHT1), APC-anti-NKG2D (eBioscience, San Diego, USA, clone CX5), APC-anti-NKp46 (eBioscience). Isotype control IgG1-FITC (DAKO), IgG1-PE (DAKO) and IgG1-APC (eBioscience) served as negative controls. NK cells were gated based on positive CD56 expression and lack of CD3 expression. At least 2,500 gated NK cells were analyzed in each reading.
Igg1 pe
Manufactured by Agilent Technologies
IgG1-PE is a fluorescent labeling reagent used in flow cytometry applications. It consists of a phycoerythrin (PE) fluorescent dye conjugated to an IgG1 antibody isotype. This product can be used to label and detect target cells or proteins of interest in samples.
Automatically generated - may contain errors
Lab products found in correlation
Igg1 fitc, by Agilent Technologies (2 mentions)
Anti cd3 fitc, by Agilent Technologies (1 mentions)
Apc anti nkg2d, by Thermo Fisher Scientific (1 mentions)
Apc igg1, by Thermo Fisher Scientific (1 mentions)
Facscalibur instrument, by BD (1 mentions)
Cd31 fitc, by Thermo Fisher Scientific (1 mentions)
Fluorescein isothiocyanate (fitc), by Thermo Fisher Scientific (1 mentions)
Cd41 pe, by Agilent Technologies (1 mentions)
Cd11b pe, by BD (1 mentions)
2 protocols using igg1 pe
1
Immunophenotyping of NK Cells
2
Flow Cytometric Analysis of Microparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
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All reagents and antibodies were diluted in a calcium chloride/PBS solution (CaCl2 2.5 mmol/L, pH 7.4, 0.2 μm filtered). The optimal antibody concentration was determined by titration. The following reagents and antibodies were used: Annexin V-Cy5-APC (Allophycocyanin, MBL international Corporation, detection of phosphatidylserine, 1:300), CD31-FITC (Fluorescein isothiocyanate, eBioscience, monoclonal mouse anti-human, 1:1000), CD41-PE (Phycoerythrin, Dako, monoclonal mouse anti-human, 1:200) and CD11b-PE (BD Biosciences, USA, monoclonal mouse anti-human, 1:100). Isotype controls used were IgG1-PE, IgG2a-PE and IgG1-FITC (all from DAKO). For the Annexin V-APC negative control, Annexin V-APC was diluted 1:300 in citrate/PBS solution. Endothelial MP were defined as CD31+ (CD41-), platelet as CD41+ (showing weak CD31+) and leukocyte MP as CD11b+.
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