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Cd16 pacific blue

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The CD16-Pacific Blue is a laboratory equipment product. It is used for the detection and analysis of CD16 surface markers on cells. The CD16-Pacific Blue provides a specific and quantitative measurement of CD16 expression levels in cell samples.

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Lab products found in correlation

Hla dr pe, by Thermo Fisher Scientific (4 mentions) Cd14 apc cy7, by BD (3 mentions) Facscanto 2, by BD (3 mentions) Cd19 fitc, by BD (2 mentions) Cd11b apc, by BD (2 mentions) Lysis fix solution, by BD (2 mentions) Cd62l apc, by BD (2 mentions) Canto 2, by BD (1 mentions) Lysis buffer, by BD (1 mentions) Cd4 pacific blue, by BD (1 mentions) Cd56 pe, by BD (1 mentions) Cd8 pe cy7, by BD (1 mentions) Nkg2a pe, by Beckman Coulter (1 mentions) Cd28 petexasred, by Beckman Coulter (1 mentions) Cd14 fitc, by BD (1 mentions) Ki67 fitc, by BD (1 mentions) Streptavidin pacific blue, by Thermo Fisher Scientific (1 mentions) Stat5 pe, by BD (1 mentions) Cd95 pe, by BD (1 mentions) Anti ccr7 150503, by R&D Systems (1 mentions)

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Anti-CD16 Pacific Blue (4 citations)

7 protocols using cd16 pacific blue

1

Neutrophil Identification by Flow Cytometry

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Circulating neutrophils were identified by flow cytometry using the protocol described in our previous publications [19 (link), 54 ]. In brief, 30 μl of fresh blood were incubated with fluorochrome-labelled antibody cocktail (CD19-FITC, CD16-Pacific Blue, CD11b-APC (all from BD Biosciences, Oxford, UK) and HLA-DR-PE (eBioscience, UK)) for 45 min in the dark at 4 °C. Red blood cells were removed with lysis buffer (BD Biosciences) and samples were acquired using a BD Canto II flow cytometer (BD Biosciences). The flow cytometry data were analysed using FlowJo (version 10, Tree Star Inc., Ashland, OR, USA). Neutrophils were identified based on their cell size (FSC) and granularity (SSC) as well as their cell surface antigens (CD11b+CD16+HLA-DR−).
Chen M., Yang N., Lechner J., Toth L., Hogg R., Silvestri G., Chakravarthy U, & Xu H. (2020). Plasma level of lipocalin 2 is increased in neovascular age-related macular degeneration patients, particularly those with macular fibrosis. Immunity & Ageing : I & A, 17, 35.
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2

Whole Blood Immunophenotyping by Flow Cytometry

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Freshly drawn blood samples (30 μl) were incubated with fluorochrome-labelled antibodies in a total volume of 100 μl FACS buffer (PBS/1% fetal calf serum (FCS)) for 45 min in the dark at 4 °C. Red blood cells were lysed by incubating the sample with 2 ml of 1X red blood cell lysis solution (BD Biosciences, Oxford, UK) for 10 min at room temperature (RT). After thorough washes samples were fixed in 1% paraformaldehyde (PFA) for 30 min in the dark at 4 °C. Samples were then washed and resuspended in PBS. All samples were examined by flow cytometry (FACS CANTO II; BD Biosciences), and data analysed blindly using the FlowJo software (version 10.07 for Windows, Tree Star, Ashland, OR, USA). The following anti-human antibodies were used for identification of leukocyte subsets: CD14-APC-Cy7, CD19-FITC, CD16-Pacific Blue, CD56-PE, CD8-PE-Cy7, CD4-Pacific Blue, CD11b-APC (all from BD Biosciences, UK) and HLA-DR-PE (eBioscience, UK). Live cells were gated for further analysis of leukocyte subsets. Different subsets of leukocytes were identified by relative cell size and granularity (FSC/SSC) or cell surface markers (Fig. 1). The neutrophil/lymphocyte ratio (NLR) was calculated by dividing the percentage of neutrophils by the percentage of lymphocytes.
Lechner J., Chen M., Hogg R.E., Toth L., Silvestri G., Chakravarthy U, & Xu H. (2015). Alterations in Circulating Immune Cells in Neovascular Age-Related Macular Degeneration. Scientific Reports, 5, 16754.
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3

Flow Cytometry Immunophenotyping Protocol

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The following antibodies were used for flow cytometry: CD3 Alexa 700 (SP34-2 BD Biosciences), CD4 AmCyan (L200 BD Biosciences), CD8 PerCP-Cy5.5 (SKI eBiosciences), CD8 AmCyan (SKI BD Biosciences), CD28 PE-Texas Red (CD28.2 Beckman Coulter, BD Biosciences), CD95 PE (DX2 BD Biosciences, eBiosciences), CCR5 Allophycocyanin (3A9 BD Biosciences), Ki-67 FITC (B56 BD Biosciences), CD56 PerCP-Cy5.5 (MEM-188 Invitrogen), CD16 Pacific Blue (3G8 BD Biosciences, Biolegend), CD20 Allophycocyanin-Cy7 (L27 BD Biosciences), HLA-DR PE-Texas Red (TU36 Invitrogen, Immu357 Beckman Coulter), NKG2A PE (Z199 Beckman Coulter), CD14 FITC (M5E2 BD Biosciences, R&D Systems), STAT5 PE (47/Stat5 (pY6) BD Biosciences), BrdU FITC (B44 BD Biosciences), BrdU Allophycocyanin (B44 BD Biosciences). Anti-CCR7 (150503) was purchased as purified immunoglobulin from R&D Systems, conjugated to biotin using a Pierce Chemical Co. biotinylation kit, and visualized with streptavidin–Pacific Blue (Invitrogen). Rhesus recombinant anti-IL-15 and rhesus recombinant control IgG1 mAb were provided through the National Institutes of Health’s Nonhuman Primate Reagent Resource Program. Simian recombinant IL-7 was provided by Cytheris SA (Issy-Les-Moulineaux, France). Rhesus recombinant IL-15 was provided by Francois Villinger (Emory University) through the Resource for Nonhuman Primate Immune Reagents.
DeGottardi M.Q., Okoye A.A., Vaidya M., Talla A., Konfe A.L., Reyes M.D., Clock J.A., Duell D.M., Legasse A.W., Sabnis A., Park B.S., Axthelm M.K., Estes J.D., Reinmann K.A., Sekaly R.P, & Picker L.J. (2016). Effect of anti-IL-15 administration on T cell and NK cell homeostasis in rhesus macaques. Journal of immunology (Baltimore, Md. : 1950), 197(4), 1183-1198.
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4

Isolation and Characterization of MCL Subpopulations

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MCL tumor cell-enriched buffy coats were isolated from apheresis or leukemic phase blood of MCL patients by Histopaque-1077 (Sigma-Aldrich, St. Louis, MO) gradient centrifugation. Obtained cells were then stained with antibodies against CD34-APC (Cat No. 555824), CD3-APC-Cy7 (Cat No. 557832), CD45-FITC (Cat No. 555482), CD19-PE (Cat No. 555413), and Sytox blue for selection of live cells (all from BD Bioscience, San Jose, CA). Subpopulations of MCL-ICs (CD34-CD3-CD45+CD19-) and MCL-non-ICs (CD34-CD3-CD45+CD19+) were isolated using a fluorescence-activated cell sorter (Influx, BD Bioscience, San Jose, CA) according to a previously described protocol [25 (link)]. Subpopulations of sorted cells were analyzed for purity by immunostaining with markers for plasma cells (CD27, CD38) and natural killer cells (CD56, CD16) using the antibodies CD27-PerCP-Cy5.5 (Cat No. 560612), CD38-PE-Cy7 (Cat No. 560677), CD56-PE-Cy7 (Cat No. 557747), and CD16-Pacific blue (Cat No. 558122) (all from BD Bioscience, San Jose, CA), respectively.
Mathur R., Sehgal L., Braun F.K., Berkova Z., Romaguerra J., Wang M., Rodriguez M.A., Fayad L., Neelapu S.S, & Samaniego F. (2015). Targeting Wnt pathway in mantle cell lymphoma-initiating cells. Journal of Hematology & Oncology, 8, 63.
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5

Flow Cytometry Immunophenotyping of Blood

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Blood samples (30 µl) were incubated with fluorochrome-labelled antibodies in a total volume of 100 µl FACS buffer (PBS/1% fetal calf serum (FCS)) for 45min. Red blood cells were removed and samples fixed with lysis/fix solution (BD Biosciences, Oxford, UK). All samples were examined by flow cytometry (FACS CANTO II; BD Biosciences), and data analysed using the FlowJo software (Tree Star, Ashland, OR, USA). The following antibodies were used: CD14-APC-Cy7, CD16-Pacific Blue, CD62L-APC, MHC-II (HLA-DR, DP, DQ)-FITC (BD Biosciences), CCR2-PerCP, CX3CR1-PE-Cy7 (BioLegend UK Ltd., London, UK) and HLA-DR-PE (eBioscience, Hatfield, UK).
Chen M., Lechner J., Zhao J., Toth L., Hogg R., Silvestri G., Kissenpfennig A., Chakravarthy U, & Xu H. (2016). STAT3 Activation in Circulating Monocytes Contributes to Neovascular Age-Related Macular Degeneration. Current Molecular Medicine, 16(4), 412-423.
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6

Intracellular Cytokine Staining Workflow

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Intracellular cytokine staining was performed as previously described(1) using the following markers: CD3-APCH7, CD107a PECy7, CD14-Pacific Blue, CD16-Pacific Blue, CD4-PECy5.5, IFN-γ-PerCPCy5.5, CD45RO-AF700, CD19-Pacific Blue (BD Biosciences, San Jose, CA), TNFα-AF647, CD8-BV570, CCR7-BV711 (BioLegend, San Diego, CA), granzyme B-PE Texas Red (Invitrogen), CD27-PECy5 (eBioscience, San Diego, CA) perforin-FITC(Abcam, Cambridge, UK). Prepared cells were acquired using an LSR II flow cytometer equipped with BD FACSDiva software (BD Biosciences). Acquired data was analyzed using the FlowJo software version 7.6.3 (Tree Star).
Morrow M.P., Kraynyak K.A., Sylvester A.J., Shen X., Amante D., Sakata L., Parker L., Yan J., Boyer J., Roh C., Humeau L., Khan A.S., Broderick K., Marcozzi-Pierce K., Giffear M., Lee J., Trimble C.L., Kim J.J., Sardesai N.Y., Weiner D.B, & Bagarazzi M.L. (2016). Augmentation of cellular and humoral immune responses to HPV16 and HPV18 E6 and E7 antigens by VGX-3100. Molecular Therapy Oncolytics, 3, 16025-.
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7

Multiparameter Flow Cytometry of Immune Cells

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Blood samples (30 µl) were incubated with fluorochrome-labelled antibodies in a total volume of 100 µl FACS buffer (PBS/1% fetal calf serum (FCS)) for 45min. Red blood cells were removed and samples fixed with lysis/fix solution (BD Biosciences, Oxford, UK). All samples were examined by flow cytometry (FACS CANTO II; BD Biosciences), and data analysed using the FlowJo software (Tree Star, Ashland, OR, USA). The following antibodies were used: CD14-APC-Cy7, CD16-Pacific Blue, CD62L-APC, MHC-II (HLA-DR, DP, DQ)-FITC (BD Biosciences), CCR2-PerCP, CX3CR1-PE-Cy7 (BioLegend UK Ltd, London, UK) and HLA-DR-PE (eBioscience, Hatfield, UK).
Chen M., Lechner J., Zhao J., Toth L., Hogg R., Silvestri G., Kissenpfennig A., Chakravarthy U, & Xu H. (2016). STAT3 Activation in Circulating Monocytes Contributes to Neovascular Age-Related Macular Degeneration. Current Molecular Medicine, 16(4), 412-423.
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