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Cd57 apc

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The CD57 APC is a laboratory reagent used for the detection and identification of cells expressing the CD57 antigen. It is a monoclonal antibody conjugated with the fluorochrome Allophycocyanin (APC).

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Lab products found in correlation

Facscanto 2, by BD (2 mentions) Hla dr fitc, by BD (2 mentions) Ccr7 pe, by BD (2 mentions) Cd27 percp cy5, by BD (2 mentions) Cd3 fitc, by BD (2 mentions) Cd3 v500, by BD (1 mentions) Cd38 pe, by BD (1 mentions) Cd4 pe cy7, by BD (1 mentions) Cd45ra pe cy7, by BD (1 mentions) Pd 1 pe, by Thermo Fisher Scientific (1 mentions) Cd4 apc efluor 780, by Thermo Fisher Scientific (1 mentions) Cd27 apc efluor780, by Thermo Fisher Scientific (1 mentions) Cd8 pacific blue, by BD (1 mentions) Cd28 percp cy5, by BD (1 mentions) Facsdiva software, by BD (1 mentions) Cd8 bv510, by BD (1 mentions) Brilliant staining buffer, by BD (1 mentions) Cd4 buv395, by BD (1 mentions) Cd27 bv421, by BD (1 mentions) Golgiplug, by BD (1 mentions)

Spelling variants of this product

Anti-CD57-APC (5 citations) CD57 (APC/clone NK-1) (2 citations)

5 protocols using cd57 apc

1

Cryopreserved PBMC Immune Phenotyping

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Cryopreserved PBMCs were used for immune phenotyping. Peripheral blood mononuclear cells were thawed and stained with monoclonal antibodies (mAbs) (30 minutes at 4°C in the dark). The following directly conjugated mAbs were used for cell surface marker staining: CD3 V500, CD4 PE-Cy7, CD8 Pacific Blue, CD45RA PE-Cy7, CCR7 PE, HLA-DR FITC, CD38 PE, CD27 PerCP Cy5.5, CD28 PerCP Cy5.5, CD57 APC (BD Biosiences, San Jose, CA), CD4 APC eFluor780, CD27 APC eFluor780, and PD-1 PE (eBioscience, San Diego, CA). Fluorescence was measured with the FACS Canto II (BD Biosciences). The proportion of T cells expressing each marker and the mean fluorescence intensity were determined using FlowJo 7.6 (TreeStar, Ashland, OR).
Kruize Z., Maurer I., van Dort K.A., van den Elshout M.A., Hoornenborg E., Booiman T., Prins M, & Kootstra N.A. (2020). Human Immunodeficiency Virus-Negative Men Who Have Sex With Men Have an Altered T-Cell Phenotype and Bioenergy Metabolism. Open Forum Infectious Diseases, 7(8), ofaa284.
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2

Phenotyping of T cell subsets

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T cell phenotyping was performed on fresh peripheral blood mononuclear cells (PBMCs) that were isolated by density gradient. Cells were resuspended in Brilliant Staining Buffer (BD Biosciences, 563794) and then stained with Fixable Viability Stain-PE (BD Biosciences, New Jersey, US, 564996) to exclude non-viable cells, and the following uorescently-conjugated monoclonal antibodies: CD3-FITC (BD Biosciences, 555332), CD4-BUV395 (BD Biosciences, 564724), CD8-BV510 (BD Biosciences, 563256), CD45RA-APC-CyTM 7 (BD Biosciences, 560674), CD27-BV421 (BD Biosciences, 562513), CD28-PE-CyTM 7 (BD Biosciences, 560684) and CD57-APC (BD Biosciences 560845). After incubation and washing, cells were analyzed by ow cytometry using BD LSR FortessaTM X-20 ow cytometer and BD FACSDivaTM software (BD Biosciences). Data were analyzed with FlowJoTM v10 software (TreeStar).
After analysis (Supplemental le 1) CD4 + and CD8 + T cell populations were identi ed as part of different phenotypes: CD8 + and CD4 + naïve T cells (TN) (CD45RA + CD27 + CD28 + CD57-), CD8 + and CD4 + TCM (CD45RA-CD27 + CD28 + CD57-); CD8 + and CD4 + TTM (CD45RA-CD27 + CD28-CD57-); CD8 + TEM (CD45RA-CD27-CD28+/-CD57-), CD4 + TEM TH1 (CD45RA-CD27-CD28-CD57-), CD4 + TEM TH2 (CD45RA-CD27-CD28 + CD57-), and CD8 + and CD4 + TEMRA (CD45RA + CD27-CD28-CD57+), as previously described 17, 18, 19, 20, 21, 22, 23, 24, 25 (Supplemental le 2).
Martínez-Román P., Cortegano I., Millares D.V., Crespo‐Bermejo C., Rallón N., Benito J.M., Lara‐Aguilar V., Arca-Lafuente S., Rickenbach M., Domínguez L., Ryan P., Santos I.d.l., Coiras M., Palladino C., Gaspar M., Fernández‐Rodríguez A., & Briz V. (2022). T cell maturation and senescence after HCV cure in HIV-HCV coinfected patients: a prospective study.
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3

Phenotypic Characterization of Activated T Cells

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Lymphocytes 5 × 106/mL were stimulated with 1 μg/mL PepMix in a CTL medium for 2 h and then for an additional 12 h in the presence of 1:1000 diluted Golgi plug (BD Biosciences, Franklin Lakes, NJ, USA). After incubation, cells were harvested with PBS and stained with the LIVE/DEAD Blue Dead Cell Staining Kit (Thermofisher) and antibodies to CD45RA-BB515, CD27-BV650, CD45RO-BV786, PD1-PECF594, CCR7-BV605 (BD Horizon, BD Biosciences), CD56-BV510, TIGIT (VSTM3)-PE/Cy7 (Biolegend, San Diego, CA, USA), CD8-AlexaFluor700 (Exbio, Prague, Czech Republic) and CD57—APC (BD Pharmingen). The cells were then washed with PBS, fixed using IC fixation and permeabilisation Buffer (eBioscience, San Diego, CA, USA) for 20 min and stained intracellularly with antibodies against IFN-gamma-PE, CD3-APCCy7 (Biolegend, San Diego, CA, USA) and CD4-PacificBlue (Exbio) in a permeabilisation buffer. The cells were washed and resuspended in a FACS buffer (FB-PBS containing 0.09% sodium azide, 1% BSA). The cells were measured using the BD LSR Fortessa 5 L flow cytometer (BD Biosciences). The obtained data were analysed by the FlowJo 10.5 software (TreeStar, Ashland, OR, USA). The gating strategy is shown in Figure 1.
Šťastná-Marková M., Hamšíková E., Hainz P., Hubáček P., Kroutilová M., Kryštofová J., Ludvíková V., Musil J., Pecherková P., Saláková M., Šroller V., Vydra J, & Němečková Š. (2021). Pretransplant BK Virus-Specific T-Cell-Mediated Immunity and Serotype Specific Antibodies May Have Utility in Identifying Patients at Risk of BK Virus-Associated Haemorrhagic Cystitis after Allogeneic HSCT. Vaccines, 9(11), 1226.
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4

Comprehensive Leukocyte Subset Analysis

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For NK and K562 cell culture, we used full media containing RPMI 1640 media (Gibco®) supplemented with 10% FBS, MEM non-essential amino acids (1%), sodium pyruvate (1%), l-glutamine (1%; all from PanBiotech), and penicillin/streptomycin (1%; Thermo Fischer Scientific). For the washing steps, we used Dulbecco’s phosphate-buffered saline (DPBS; Gibco®).
To analyze the different leukocyte subsets, CD3, CD14, CD16, CD19, CD45, and CD56 antibodies with different fluorochromes from Becton Dickinson (BD) were used. For detailed NK cell subset analyses, we used anti-KIR2D-, KIR3DL1/2-, and KIR2DL1/S1-PE (Miltenyi), KIR2DL1-PerCP (R&D), KIR2DL2/3/S2-APC (Beckman Coulter), KIR3DL1 PE-Vio770 (Miltenyi), NKG2A FITC (Miltenyi), and CD57 APC (BD). For the KIR staining, the clones of the antibodies were selected according to Czaja et al. (17 (link)), and a sequential staining protocol was used as described by Beziat et al. (18 (link)). For intracellular staining, we used IFN-γ PE-Cy-7 and MIP-1β APC-H7 (BD). To exclude dead cells, 7-AAD (BD) for extracellular and Fixable Viability Dye eFluor® 520 (eBioscience) for intracellular staining were used.
Jacobs B., Tognarelli S., Poller K., Bader P., Mackensen A, & Ullrich E. (2015). NK Cell Subgroups, Phenotype, and Functions After Autologous Stem Cell Transplantation. Frontiers in Immunology, 6, 583.
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5

Phenotyping of Immune Cells by Flow Cytometry

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Antibodies for surface markers staining of DCs and T cells were obtained from BD Biosciences, New York, USA (anti-human CD3-PE, CD3-FITC, CD8-PerCP, CD8-APC, CD56-PE, NKG2D-APC, CD4-FITC, CD4-PerCP, CD107a-FITC, CD25-APC, CD45RO-FITC, CD27-PerCPCY5.5, CD57-APC, CCR7-PE, CD14-APC, CD80-PE, CD83-APC, CD86-FITC, HLA-DR-FITC). Antibodies for intracellular proteins staining were also obtained from BD Biosciences (anti-human IFNγ-APC, TNFα-PECY7, granzyme B-FITC, FoxP3-PE). The intracellular staining was performed by fixing and permeabilizing cells with Cytofix/Cytoperm (BD Biosciences). The experiments were performed by using FACS CantoII (BD Biosciences) flow cytometer, and data were analyzed by using the Flowjo software.
Han Y., Wu Y., Yang C., Huang J., Guo Y., Liu L., Chen P., Wu D., Liu J., Li J., Zhou X, & Hou J. (2017). Dynamic and specific immune responses against multiple tumor antigens were elicited in patients with hepatocellular carcinoma after cell-based immunotherapy. Journal of Translational Medicine, 15, 64.
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