Cd27 pe cy5 1a4cd27

Manufactured by Beckman Coulter

Sourced in United States

The CD27 PE-Cy5 (1A4CD27) is a fluorescent-labeled antibody used in flow cytometry applications. It binds to the CD27 antigen, which is expressed on various immune cells. This product can be used to identify and analyze CD27-positive cell populations in research samples.

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Lab products found in correlation

Transcription factor perm wash buffer, by Thermo Fisher Scientific (6 mentions) Ifnγ bv711 4s b3, by BioLegend (6 mentions) Cd4 bv785 okt4, by BioLegend (5 mentions) Cd3 bv650 okt3, by BioLegend (5 mentions) Cd8 bv510 rpa t8, by BioLegend (4 mentions) Lsr 2, by BD (4 mentions) Il 2 pe dazzle mq1 17h12, by BioLegend (4 mentions) Hla dr bv605 l243, by BioLegend (4 mentions) Klrg1 percp efluor 710 13f12f2, by Thermo Fisher Scientific (3 mentions) Cd153, by R&D Systems (3 mentions) Tnfα efluor 450 mab11, by BioLegend (3 mentions) Mip 1β alexa fluor 488, by R&D Systems (2 mentions) Hla dr bv605, by BioLegend (2 mentions) Eomes efluor 660 wd1928, by Thermo Fisher Scientific (2 mentions) Ki67 percpcy5.5 b56, by BD (2 mentions) Tnf α pe cy7 mab11, by BioLegend (2 mentions) Cd38 apc hit2, by BD (2 mentions) Cd19 bv750 hib19, by BioLegend (2 mentions) Cd4 bv785, by BioLegend (1 mentions) Cd3 bv650, by BioLegend (1 mentions)

Close spelling variants of this product

CD27-PE/Cy5 (3 citations) CD27-PE (3 citations)

7 protocols using cd27 pe cy5 1a4cd27

Multicolor Cytokine Analysis of Mtb300-specific CD4 T cells

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Cryopreserved cells were thawed, washed and permeabilised with a Transcription Factor perm/wash buffer (eBioscience). Cells were then stained at room temperature for 45 min with the following antibodies: CD3 BV650 (OKT3; BioLegend, San Diego, CA, USA), CD4 BV785 (OKT4; BioLegend), CD8 BV510 (RPA‐T8; BioLegend), CD27 PE‐Cy5 (1A4CD27; Beckman Coulter, Brea), HLA‐DR BV605 (L243; BioLegend), Killer cell Lectin‐like Receptor G1 (KLRG1) PerCP‐eFluor 710 (13F12F2; eBioscience), IFN‐γ BV711 (4S.B3; BioLegend), TNF‐α eFluor 450 (Mab11; BioLegend eBioscience), IL‐2 PE/Dazzle (MQ1‐17H12, BioLegend eBioscience) MIP‐1β Alexa Fluor 488 (#24006; R&D systems, Minneapolis, MN, USA) and CD153 (R&D116614; R&D systems). Samples were acquired on a BD LSR‐II and analysed using FlowJo (v9.9.6, FlowJo LCC, Ashland, OR, USA). A positive cytokine response was defined as at least twice the background of unstimulated cells. To define the phenotype of Mtb300‐specific CD4 T cells, a cut‐off of 30 events was used.

Riou C., Du Bruyn E., Ruzive S., Goliath R.T., Lindestam Arlehamn C.S., Sette A., Sher A., Barber D.L, & Wilkinson R.J. (2020). Disease extent and anti‐tubercular treatment response correlates with Mycobacterium tuberculosis‐specific CD4 T‐cell phenotype regardless of HIV‐1 status. Clinical & Translational Immunology, 9(9), e1176.

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Phenotypic Analysis of Mycobacterial-Specific T Cells

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Cryopreserved cells were thawed, washed and permeabilized with a Transcription Factor perm/wash buffer (eBioscience). Cells were then stained at room temperature for 45 minutes with the following antibodies: CD3 BV650 (OKT3; Biolegend), CD4 BV785 (OKT4; Biolegend), CD8 BV510 (RPA-T8; Biolegend), CD27 PE-Cy5 (1A4CD27; Beckman Coulter), HLA-DR BV605 (L243; Biolegend), Killer cell Lectin-like Receptor G1 (KLRG1) PerCP-eFluor 710 (13F12F2, eBioscience), Eomes eFluor 660 (WD1928, eBioscience), IFNγ BV711 (4S.B3; Biolegend), TNFα eFluor 450 (Mab11; Biolegend), IL-2 PE/Dazzle (MQ1-17H12, Biolegend) and CD153 (R&D116614, R&D). Samples were acquired on a BD LSR-II and analyzed using FlowJo (v9.9.6, TreeStar). A positive cytokine response was defined as at least twice the background of unstimulated cells. To define the phenotype of Mtb300-specific cells, a cut-off of 30 events was used. The gating strategy is presented in Supplementary Fig. 5.

Du Bruyn E., Ruzive S., Lindestam Arlehamn C.S., Sette A., Sher A., Barber D.L., Wilkinson R.J, & Riou C. (2020). Mycobacterium tuberculosis-specific CD4 T cells expressing CD153 inversely associate with bacterial load and disease severity in human tuberculosis. Mucosal Immunology, 14(2), 491-499.

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Multiparameter Flow Cytometry for B Cell Analysis

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The following antibodies were used in three different staining panels: CD19 ECD (J3-119), CD27 PE-Cy5 (1A4CD27; both Beckman Coulter), IgD APC-Cy7 (IA6-2), CD10 BV605 (HI10a), CD21 PE-Cy7 (Bu32), CD40 PerCP-Cy5.5 (5C3; all Biolegend), CD38 APC (HIT2), CD86 PE (IT2.2), CD3 PE-Cy7 (SK7), HLA-DR APC-Cy7 (L243; all BD Biosciences), CD4 PE-Cy5.5 (S3.5), CD8 Qdot-705 (3B5), CD19 Pacific Blue (SJ25-CI), CD14 Pacific Blue (Tük4), CD3 Pacific Blue (UCHT1), Ki67 FITC (7B11; all Invitrogen) and a violet viability reactive dye (“Vivid”; Molecular Probes). All antibodies were titrated prior to use to obtain optimal titers for staining. Briefly, PBMC were stained with Vivid, then labeled with antibodies against surface markers, fixed, permeabilized and subsequently stained intracellularly with Ki67. Cells were then re-suspended in 1X CellFix (BD Biosciences) and kept at 4⁰C until acquisition. Samples were acquired on a BD Fortessa using FACSDiva software and analyzed using FlowJo (version 9.9.3; TreeStar). The gating strategy to identify B cell subsets is shown in Supplemental Figure S1.

Tanko R.F., Soares A.P., Müller T.L., Garrett N.J., Samsunder N., Abdool Karim Q., Abdool Karim S.S., Riou C, & Burgers W.A. (2016). Effect of antiretroviral therapy on the memory and activation profiles of B cells in HIV-infected African women. Journal of immunology (Baltimore, Md. : 1950), 198(3), 1220-1228.

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Multiparametric Phenotyping of Mtb300-Specific T Cells

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Cryopreserved cells were thawed, washed and permeabilized with a Transcription Factor perm/wash buffer (eBioscience). Cells were then stained at room temperature for 45 minutes with the following antibodies: CD3 BV650 (OKT3; Biolegend, San Diego, CA, USA), CD4 BV785 (OKT4; Biolegend), CD8 BV510 (RPA-T8; Biolegend), CD27 PE-Cy5 (1A4CD27; Beckman Coulter, Brea, CA, USA), HLA-DR BV605 (L243; Biolegend), Ki67 PerCPcy5.5. (B56, BD), Granzyme B (GrB) BV421 (GB11, BD), Killer cell Lectin-like Receptor G1 (KLRG1) APC (13F12F2, eBioscience), IFN-γ BV711 (4S.B3; Biolegend), TNF-α PEcy7 (Mab11; Biolegend), IL-2 PE/Dazzle (MQ1-17H12, Biolegend), Mip-1β Alexa Fluor 488 (#24006, R&D systems, Minneapolis, MN, USA) and CD153 (R&D116614, R&D). Samples were acquired on a BD LSR-II and analyzed using FlowJo (v9.9.6, FlowJo LCC, Ashland, OR, USA). The granulocytes/monocytes population was defined based on their FSC/SSC characteristics. A positive cytokine response was defined as at least twice the background of unstimulated cells. To define the phenotype of Mtb300-specific cells, a cut-off of 30 events was used.

Du Bruyn E., Ruzive S., Howlett P., Cerrone M., Jacobs A.J., Arlehamn C.S., Sette A., Sher A., Mayer-Barber K.D., Barber D.L., Mayosi B., Ntsekhe M., Wilkinson R.J, & Riou C. (2022). Comparison of the frequency and phenotypic profile of Mycobacterium tuberculosis-specific CD4 T cells between the site of disease and blood in pericardial tuberculosis. Frontiers in Immunology, 13, 1009016.

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Multiparametric Flow Cytometry Profiling

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Cell staining was performed on cryopreserved cells that were thawed, washed and permeabilized with a Transcription Factor perm/wash buffer (eBioscience). Cells were then stained at room temperature for 45 min with antibodies for CD3 BV650 (OKT3, Biolegend), CD4 BV785 (OKT4, Biolegend), CD8 BV510 (RPA-8, Biolegend), CD19-BV750 (HIB19, Biolegend), CD45RA Alexa 488 (HI100, Biolegend), CD27 PE-Cy5 (1A4CD27, Beckman Coulter), CD38 APC (HIT2, BD Biosciences), HLA‐DR BV605 (L243, Biolegend), Ki67 PerCP-Cy5.5 (B56, BD Biosciences), PD-1 PE (J105, eBioscience), Granzyme B (GrB) BV421 (BG11, BD Biosciences), IFNγ BV711 (4S.B3, Biolegend), TNFα PE-Cy7 (MAB11, Biolegend) and IL-2, PE/Dazzle 594 (MQ1-17H12, Biolegend). Samples were acquired on a BD LSR-II and analyzed using FlowJo (v9.9.6, FlowJo LCC, Ashland, OR, USA). A positive response was defined as any cytokine response that was at least twice the background of unstimulated cells. To define the phenotype of SARS-CoV-2-specific CD4 T cells, a cut-off of 20 events was used.

Riou C., du Bruyn E., Stek C., Daroowala R., Goliath R.T., Abrahams F., Said-Hartley Q., Allwood B.W., Hsiao N.Y., Wilkinson K.A., Arlehamn C.S., Sette A., Wasserman S, & Wilkinson R.J. (2021). Relationship of SARS-CoV-2–specific CD4 response to COVID-19 severity and impact of HIV-1 and tuberculosis coinfection. The Journal of Clinical Investigation, 131(12), e149125.

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Phenotypic Analysis of Mtb300-specific T Cells

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Cryopreserved cells were thawed, washed and permeabilized with a Transcription Factor perm/wash buffer (eBioscience). Cells were then stained at room temperature for 45 min with the following antibodies: CD3 BV650 (OKT3; Biolegend), CD4 BV785 (OKT4; Biolegend), CD8 BV510 (RPA-T8; Biolegend), CD27 PE-Cy5 (1A4CD27; Beckman Coulter), HLA-DR BV605 (L243; Biolegend), Killer cell Lectin-like Receptor G1 (KLRG1) PerCP-eFluor 710 (13F12F2, eBioscience), Eomes eFluor 660 (WD1928, eBioscience), IFNγ BV711 (4S.B3; Biolegend), TNFα eFluor 450 (Mab11; Biolegend), IL-2 PE/Dazzle (MQ1-17H12, Biolegend), and CD153 (R&D116614, R&D). Samples were acquired on a BD LSR-II and analyzed using FlowJo (v9.9.6, TreeStar). A positive cytokine response was defined as at least twice the background of unstimulated cells. To define the phenotype of Mtb300-specific cells, a cut-off of 30 events was used. The gating strategy is presented in Supplementary Fig. 5.

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Phenotyping SARS-CoV-2-specific T cells

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Cell staining was performed on cryopreserved cells that were thawed, washed, and permeabilized with a transcription factor perm/wash buffer (eBioscience, Thermo Fisher Scientific). Cells were then stained at room temperature for 45 minutes with antibodies for CD3 BV650 (OKT3, BioLegend), CD4 BV785 (OKT4, BioLegend), CD8 BV510 (RPA-8, BioLegend), CD19-BV750 (HIB19, BioLegend), CD45RA Alexa Fluor 488 (HI100, BioLegend), CD27 PE-Cy5 (1A4CD27, Beckman Coulter), CD38 APC (HIT2, BD Biosciences), HLA-DR BV605 (L243, BioLegend), Ki67 PerCP-Cy5.5 (B56, BD Biosciences), PD-1 PE (J105, eBioscience, Thermo Fisher Scientific), GrB BV421 (BG11, BD Biosciences), IFN-γ BV711 (4S.B3, BioLegend), TNF-α PE-Cy7 (MAB11, BioLegend), and IL-2 PE/Dazzle 594 (MQ1-17H12, BioLegend). Samples were acquired on a BD Biosciences LSR II and analyzed using FlowJo v9.9.6. A positive response was defined as any cytokine response that was at least twice the background of unstimulated cells. To define the phenotype of SARS-CoV-2–specific CD4⁺ T cells, a cutoff of 20 events was used.

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