Cd28 percp cy5

Manufactured by BD

CD28-PerCP-Cy5.5 is a fluorescent-conjugated antibody that binds to the CD28 cell surface receptor. It is designed for use in flow cytometry applications to identify and quantify CD28-expressing cells.

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Lab products found in correlation

Cd45ra pe cy7, by BD (3 mentions) Facscanto 2, by BD (2 mentions) Cd27 percp cy5, by BD (2 mentions) Ccr7 pe cy7, by BD (2 mentions) Cd8 apc h7, by BD (2 mentions) Cd3 v500, by BD (1 mentions) Hla dr fitc, by BD (1 mentions) Cd38 pe, by BD (1 mentions) Cd4 pe cy7, by BD (1 mentions) Ccr7 pe, 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) Cd57 apc, by BD (1 mentions) Cd4 alexafluor700, by BD (1 mentions) Lsr 2 cytometer, by BD (1 mentions) Cd27 apc, by BD (1 mentions) Cd45ra pacificblue, by Thermo Fisher Scientific (1 mentions) Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

PerCP-Cy5.5 (120 citations)

6 protocols using cd28 percp cy5

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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Multiparametric Flow Cytometry Immunophenotyping

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Blood samples were analysed by 8-colour flow-cytometry assays, which were performed similarly as previously described.^{2 (link)} Shortly, frozen PBMC aliquots were quickly thawed, and washed using an automatic cell washer. Following primary wash step, cell viability and cell numbers were analyzed using Vi cell counter. PBMC staining was performed with CD3-QDot605 and CD45RA-PacificBlue (both Invitrogen), CD4-AlexaFluor700, CD8-APC-H7, CCR7-PE/Cy7, CD27-APC, and CD28-PerCp/Cy5.5 (all BD Biosciences). Following three wash steps, LIVE/DEAD® Fixable Aqua Dead Cell Stain (Thermo Fisher Scientific, Waltham, MA, USA) was added to allow discrimination of viable and dead cells. Samples were measured on a BD LSR II cytometer using BD FACSDiva acquisition software. At least 100,000 viable cell events per sample were acquired. The gating scheme is depicted in Fig. 2a.

Martin-Ruiz C., Hoffmann J., Shmeleva E., Zglinicki T.V., Richardson G., Draganova L., Redgrave R., Collerton J., Arthur H., Keavney B, & Spyridopoulos I. (2020). CMV-independent increase in CD27−CD28+ CD8+ EMRA T cells is inversely related to mortality in octogenarians. NPJ Aging and Mechanisms of Disease, 6, 3.

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Flow Cytometry Analysis of Bone Marrow and Peripheral Blood Cells

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For the flow cytometry analysis, EDTA anticoagulated samples were used. The bone marrow samples were strained using special tubes (5 ml polystyrene round-bottom tube with cell-strainer cap, cat.no:352235, BD Biosciences). For the staining of the cells (BM-MNC or PBMC), the following markers were used, according to the manufacturers protocol: CD19 PC7 (cat. no.: BCI-F-IM3628, Beckman Coulter Company), CD20 APC-H7, (cat.no.: 561172, BD Biosciences), CD27 PerCP-Cy5.5 (cat.no.: 560612, BD Biosciences), CD28, PerCP-Cy5.5 (cat.no.: 337181, BD Biosciences), CD38 APC (cat.no.: 345807, BD Biosciences), CD45 Krome Orange (cat.no.: BCI-F-IM36294, Beckman Coulter Company), CD56 FITC (cat. no:345811, BD Biosciences), CD81 APC-H7 (cat. no:656647, BD Biosciences), CD117 PE, (cat. no.: 332785, BD Biosciences), CD138 VioBlue (cat.no.:130-119-843, Miltenyi Biotec), Kappa FITC (cat. no.:349516, BD Biosciences) (intracellular staining), Lambda PE (cat. no.:349516, BD Biosciences) (intracellular staining). For the intracellular staining, BD Intrasure Kit was used (cat.no.: 641778 BD Biosciences). The acquisition of the samples was done on a FACS Canto II cytometer (BD Biosciences), and the analyses were done with the Infinicyt (ver.2.0.2c, Cytognos S.L.) software.

Marx A., Osváth M., Szikora B., Pipek O., Csabai I., Nagy Á., Bödör C., Matula Z., Nagy G., Bors A., Uher F., Mikala G., Vályi-Nagy I, & Kacskovics I. (2023). Liquid biopsy-based monitoring of residual disease in multiple myeloma by analysis of the rearranged immunoglobulin genes–A feasibility study. PLOS ONE, 18(5), e0285696.

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Comprehensive CD4+ T Cell Profiling

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One day post isolation FACS analysis was used to determine the percentages of various CD4+ T cell subsets, like naïve (TN), effector memory (TEM), central memory (TCM), T helper 17 cells (Th17), follicular helper T cells (Tfh) and regulatory T cells (Treg). For this the following antibodies were used: CD4-APC H7 (BD Biosciences, 560158), CD45RA-PE Cy7 (BD Biosciences, 560675), CD45RO-PerCP Cy5.5 (BD Biosciences, 560607), CCR7-APC (BioLegend, 353214), CCR6-PE (BD Biosciences, 559562), CXCR5-PerCP Cy5.5 (BioLegend, 335001), CD127-PE Cy7 (BD Biosciences, 560822), CD25-Horizon V450 (BD Biosciences, 560355). Further, the expression of different receptors on unstimulated as well as stimulated CD4+ T cells was determined using the following antibodies: CD4-APC H7, TCR-CD3-APC H7 (BD Biosciences, 560275), CD28-PerCP Cy5.5 (BD Biosciences, 560685), MHC-I-Horizon V450 (BD Biosciences, 561346), FAS-PE (BD Biosciences, 556641), FAS-L-PE (BioLegend, 306407), PD1-APC (BD Biosciences, 558694), PD1-L-PE Cy7 (BD Biosciences, 558017), TRAIL-PE (BD Biosciences, 550516), CTLA-4-PE (BD Biosciences, 555853), CD69-Horizon V450 (BD Biosciences, 560740), CD25-PE Cy7 (BD Biosciences, 557741), CXCR4-PerCP Cy5.5 (BD Biosciences, 560670) and CCR5-PE Cy7 (BD Biosciences, 557752). Cells were analyzed using the BD FACS Canto II with FACSDiva software.

Heigele A., Joas S., Regensburger K, & Kirchhoff F. (2015). Increased susceptibility of CD4+ T cells from elderly individuals to HIV-1 infection and apoptosis is associated with reduced CD4 and enhanced CXCR4 and FAS surface expression levels. Retrovirology, 12, 86.

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Comprehensive Phenotypic Analysis of γδ T Cells

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Freshly isolated and 5-day proliferated γδ T cells were membrane-stained with the following monoclonal antibodies; γδ TCR-FITC (Miltenyi), CD56-PE (BD), CD69-PE (BD), and HLA-DR-PE (BD). Propidium iodide (PI; Life Technologies) was added to exclude dead cells from phenotypic analysis. Data acquisition was performed on a FACScan multiparametric flow cytometer (BD). Phenotypic characterization of γδ T cells was examined pre- and post-expansion, using CD27-FITC (BD), CD69-FITC (BD), CD56-PE (BD), CD80-PE (BD), CD45RA-PE-Cy7 (BD), CD28-PerCP-Cy5.5 (BD), CD16-PB (BD), CD86-V450 (BD), γδ TCR-APC (Miltenyi), and HLA-DR-APC-H7 (BD). Live/Dead^® Fixable Aqua Stain was used to distinguish viable from non-viable cells. Data were acquired on a FACSAria II flow cytometer (BD). Corresponding species- and isotype-matched antibodies were used as controls.

Van Acker H.H., Anguille S., Willemen Y., Van den Bergh J.M., Berneman Z.N., Lion E., Smits E.L, & Van Tendeloo V.F. (2016). Interleukin-15 enhances the proliferation, stimulatory phenotype, and antitumor effector functions of human gamma delta T cells. Journal of Hematology & Oncology, 9, 101.

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Comprehensive Immune Cell Profiling

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The following conjugated antibodies were used for cell-surface staining: CD3-Pacific Blue (BD Pharmingen; Clone UCHT1; Cat. No. 558117; San Diego, CA, USA), CD8-APC H7 (BD Pharmingen; Clone SK1; Cat. No. 641400), CD45RA-PE (BD Pharmingen; Clone HI100; Cat. No. 555489), CCR7-PE-Cy7 (BD Pharmingen; Clone 3D12; Cat. No. 557648), CD28-PerCP-Cy5.5 (BD Biosciences; Clone L293; Cat. No. 337181; San Jose, CA, USA), CD27-Alexa Fluor 700 (BD Pharmingen; Clone M-T271; Cat. No. 560611), CD95-APC (BD Pharmingen; Clone DX2; Cat. No. 558814) and CD127-FITC (BD Pharmingen; Clone HIL-7R-M21; Cat. No. 560549). Conjugated antibodies for intracellular staining included the following: IFN-γ-FITC (BD Pharmingen; Clone 4S.B3; Cat. No. 554551), IL-2-PerCP-Cy5.5 (BD Pharmingen; Clone MQ1-17H12; Cat. No. 560708) and TNF-α-AlexaFluor 700 (BD Pharmingen; Clone MAb11; Cat. No. 557996). To exclude dead cells, the Fixable Aqua Dead Cell Stain viability marker was used (Invitrogen; Cat. No. L34957; Eugene, OR, USA).

Mateus J., Lasso P., Pavia P., Rosas F., Roa N., Valencia-Hernández C.A., González J.M., Puerta C.J, & Cuéllar A. (2015). Low Frequency of Circulating CD8+ T Stem Cell Memory Cells in Chronic Chagasic Patients with Severe Forms of the Disease. PLoS Neglected Tropical Diseases, 9(1), e3432.

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