Cd8 pe cy7

Manufactured by Beckman Coulter

Sourced in Austria, United Kingdom

The CD8-PE-Cy7 is a fluorescent-labeled antibody used for the detection and quantification of CD8-positive cells in flow cytometry analysis. It is composed of an anti-CD8 antibody conjugated with the PE-Cy7 fluorescent dye. The CD8 antigen is expressed on the surface of cytotoxic T lymphocytes, which play a crucial role in the adaptive immune response.

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Lab products found in correlation

Ficoll hypaque, by GE Healthcare (1 mentions) Cd4 apc, by BD (1 mentions) Cd3 fitc, by BD (1 mentions) Moflo astrios cell sorter, by Beckman Coulter (1 mentions) Cd19 percp cy5, by Thermo Fisher Scientific (1 mentions) Cd56 v450, by BD (1 mentions) Cd3 ecd, by Beckman Coulter (1 mentions) Cd10 pe cy5, by Beckman Coulter (1 mentions) Cd19 pe cy5, by Beckman Coulter (1 mentions) Anti brdu fitc antibody, by BD (1 mentions) Anti cd4 fitc, by BD (1 mentions) Cd4 pe cy7, by Thermo Fisher Scientific (1 mentions) Anti cd8 pe cy7, by BioLegend (1 mentions) Live dead stain kit, by Thermo Fisher Scientific (1 mentions) Cd4 pe, by Beckman Coulter (1 mentions) Cd44 fitc, by Beckman Coulter (1 mentions) Cd62l apc, by Beckman Coulter (1 mentions) Cd3 apc h7, by BD (1 mentions) Anti cd28 cd49d antibodies, by BD (1 mentions) Ficoll paque, by GE Healthcare (1 mentions)

Spelling variants of this product

CD8-PE (7 citations) PE/Cy7 anti-human CD8 (2 citations)

9 protocols using cd8 pe cy7

Isolation and Sorting of Leukocyte Subsets

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Peripheral blood mononuclear cells (PBMCs) of both patients were isolated by density gradient centrifugation using Ficoll-Hypaque (GE Healthcare, Uppsala, Sweden) and stained with the following antibodies: CD3-FITC, CD4-APC (BD, Biosciences, Schwechat, Austria), CD8-PECy7 (Beckmann Coulter, Krefeld, Germany), CD19-PerCPCy5.5 (eBioscience, Vienna, Austria) and CD56-V450 (BD, Biosciences, Austria). Subsequently, the stained cells were sorted into different subgroups of leukocytes CD3+CD4+CD8-T-cells, CD3+CD4-CD8+ T-cells and CD3-CD19+ B-cells using a MoFlo Astrios Cell Sorter from Beckmann Coulter.

Ban S.A., Salzer E., Eibl M.M., Linder A., Geier C.B., Santos-Valente E., Garncarz W., Lion T., Ott R., Seelbach C., Boztug K, & Wolf H.M. (2014). Combined Immunodeficiency Evolving into Predominant CD4+ Lymphopenia Caused by Somatic Chimerism in JAK3. Journal of Clinical Immunology, 34(8), 941-953.

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Profiling pSTAT3 Expression in PTCL, NOS

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Flowcytometric assay was performed on three PTCL, NOS cases, using one excised tonsil case as control. All PTCL, NOS cases were diagnosed in our laboratory using a T-cell lymphoma panel recommended by Euroflow and excluded from PTCL of T follicular helper (Tfh) cell phenotype (19 (link)) on a Cytek Spectrum Flowcytometer (NL-CLC, V16-B14). The detection of pSTAT3-Y705 was by using commercially available antibody (Catalog No: 612569) from BD Pharmingen (BD Bioscience, USA) and pSTAT3-S727 by conjugating fluorescence tag (FlexAble Coralite V405 Antibod Labeling Kit For Rabbit IgG, Cat No: KFA006) with pSTAT3-S727 (ab32143, Abcam) from Abcam (Cambridge, UK), in an 8-color panel consisting of pSTAT3-Y705-FITC, pSTAT3-S727-Corolite V405, CD3-ECD (Catlog No: A07748, Beckman Coulter), CD8-PE-Cy7 (Catlog No: 6607102, Beckman Coulter), CD10-PE-Cy5 (Catlog No: A07761, Beckman Coulter), CD19-PE-Cy5.5 (Catlog No: A66328, Beckman Coulter), CD45-cFlourV547 (Catlog No: R7-10011, Cytek), CD4-cFlourV610 (Catlog No: R7-20073, Cytek) and PD-1-BV750 (Catlog No: 329966, Biolegend).

Xiang C., Wu W., Fan M., Wang Z., Feng X., Liu C., Liu J., Liu G., Xia L., Si H., Gu Y., Liu N., Luo D., Wang Y., Ma D., Hu S, & Liu H. (2023). Phosphorylated STAT3 as a potential diagnostic and predictive biomarker in ALK- ALCL vs. CD30high PTCL, NOS. Frontiers in Immunology, 14, 1132834.

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Multiparametric Flow Cytometry Analysis

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Stained cells were analyzed on an LSR cytometer (Becton Dickinson). To determine memory phenotype, CD4⁺ T cells were stained with anti -CD8-PEcy7 (Biolegend), -CD4-PE, -CD44-FITC and -CD62L-APC (Beckman Coulter) antibodies. For double negative population analysis, cells were stained with anti-CD4-FITC (PharMingen), -CD8-PEcy7, -B220-APC (Beckman Coulter) and -TCR-PE (PharMingen) antibodies.
BrdU was detected with an anti-BrdU-FITC antibody (Becton Dickinson) combined with -CD4-PEcy7 (eBioscience), -CD8-SPRD (Beckman Coulter), -CD44-biot (PharMingen), Av-SPRD (Beckman Coulter), -CD62L-PE (Southern) and -TCR-PE (PharMingen) antibodies. To distinguish live from dead cells, we preincubated cells with the LIVE/DEAD stain kit (Invitrogen) according to manufacturer's instructions. Only live cells were considered for analyses.

Daszkiewicz L., Vázquez-Mateo C., Rackov G., Ballesteros-Tato A., Weber K., Madrigal-Avilés A., Di Pilato M., Fotedar A., Fotedar R., Flores J.M., Esteban M., Martínez-A C, & Balomenos D. (2015). Distinct p21 requirements for regulating normal and self-reactive T cells through IFN-γ production. Scientific Reports, 5, 7691.

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Phospho-Signaling Pathway Analysis in Activated PBMCs

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Peripheral blood mononuclear cells (PBMCs) were separated on Ficoll-Paque (GE Healthcare) according to the manufacturer's instructions. PBMCs were reconstituted in a culture medium consisting of RPMI 1640 with 25 mM HEPES, L-glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin (Lonza, Basel, Switzerland) to a final concentration of 2 million cells per milliliter. After a 1-h rest at 37°C in a 5% CO₂ atmosphere, the cells were stimulated on 96-well plate containing coated anti-CD3 (10 μg/ml, Exbio Praha) and free costimulatory anti-CD28/CD49d antibodies (1 μg/ml, BD Biosciences) for 5, 15, and 30 min. The cells were fixed with 4% formaldehyde for 10 min and permeabilized with ice-cold methanol for 30 min. The following fluorochrome conjugates were used for cytometric detection: phospho-Akt (Ser473)-Alexa Fluor 488, phospho-S6 (Ser235/236)-Pacific Blue (Cell Signaling Technologies), phospho-mTOR (Ser2448)-PE (eBioscience, Thermo Fisher), CD45-Pacific Orange, CD45RA-APC (Exbio), CD8-PE-Cy7 (Beckman Coulter), CD4-PerCP-Cy5.5, and CD3-APC-H7 (BD Biosciences). The samples were acquired on Canto II flow cytometer and analyzed using FlowJo software (BD Biosciences).

Polaskova K., Merta T., Martincekova A., Zapletalova D., Kyr M., Mazanek P., Krenova Z., Mudry P., Jezova M., Tuma J., Skotakova J., Cervinkova I., Valik D., Zdrazilova-Dubska L., Noskova H., Pal K., Slaby O., Fabian P., Kozakova S., Neradil J., Veselska R., Kanderova V., Hrusak O., Freiberger T., Klement G.L, & Sterba J. (2020). Comprehensive Molecular Profiling for Relapsed/Refractory Pediatric Burkitt Lymphomas—Retrospective Analysis of Three Real-Life Clinical Cases—Addressing Issues on Randomization and Customization at the Bedside. Frontiers in Oncology, 9, 1531.

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Multicolor Flow Cytometry Panel

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Fluorescent tagged antibodies used were specific for: CD3 allophycocyanin (APC), CD3 APC-cyanine7 (Cy7), CD4 phycoerythrin (PE), IL5 PE (BD Biosciences, San Jose CA); CD8 PE-Cy7 (Beckman Coulter Miami FL); IFN-gamma APC, FoxP3 (clone PCH101) and its isotype control (eBiosciences, San Diego CA); CD25-PE (Miltenyi Biotec Inc, Auburn CA).

Dillon P.M., Olson W.C., Czarkowski A., Petroni G.R., Smolkin M., Grosh W.W., Chianese-Bullock K.A., Deacon D.H, & Slingluff CL J.r. (2014). A melanoma helper peptide vaccine increases Th1 cytokine production by leukocytes in peripheral blood and immunized lymph nodes. Journal for Immunotherapy of Cancer, 2, 23.

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Multiparametric Flow Cytometry Analysis

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Flow cytometric analysis was performed on the BAL samples with Becton Dickinson Biosciences FACSCanto II flow cytometer (BD, Franklin Lakes, NJ, USA). Data analysis was performed using FACSDiva software (BD). The following antibodies were used in this study: CD45 Krome Orange (Beckman Coulter, clone J33), CD3 Alexa Fluor 700 (BD, clone UCHT1), CD56 FITC (BD, clone NCAM16.2), CD19 BV421 (BD, clone HIB19), CD4 Pe-Cy5.5 (Beckman Coulter, clone 13B8.2), CD8 Pe-Cy7 (Beckman Coulter, clone SFCI21Thy2D3), CD38 APC (BD, clone HB7), HLA-DR (BD, L243), and CD25 PE (BD, clone 2A3).

Gelarden I., Nguyen J., Gao J., Chen Q., Morales-Nebreda L., Wunderink R., Li L., Chmiel J.S., Hrisinko M., Marszalek L., Momnani S., Patel P., Sumugod R., Chao Q., Jennings L.J., Zembower T.R., Ji P, & Chen Y.H. (2021). Comprehensive evaluation of bronchoalveolar lavage from patients with severe COVID-19 and correlation with clinical outcomes. Human Pathology, 113, 92-103.

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Multiparametric Analysis of T-cell Responses

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ICS was performed as reported earlier [23 (link)]. Cells were stimulated with individual peptides or with an equal volume of water/ 10% DMSO in the presence of anti-CD107a (BD), GolgiStop (BD) and Brefeldin A (Sigma-Aldrich). After 12 h cells were stained for CD4-APC-Cy7 (BD), CD8-PECy7 (Beckman Coulter) and CD3-BV711 (Biolegend) and with Aqua Live Dead, fixed and permeabilized in (Cytoperm/Cytofix; BD) and further stained for IFNγ-Alexa Fluor 700 (BD Biosciences), anti-TNF-Pacific Blue (Biolegend), IL-10-PE and IL-2-APC (both BD).

Rammensee H.G., Wiesmüller K.H., Chandran P.A., Zelba H., Rusch E., Gouttefangeas C., Kowalewski D.J., Di Marco M., Haen S.P., Walz J.S., Gloria Y.C., Bödder J., Schertel J.M., Tunger A., Müller L., Kießler M., Wehner R., Schmitz M., Jakobi M., Schneiderhan-Marra N., Klein R., Laske K., Artzner K., Backert L., Schuster H., Schwenck J., Weber A.N., Pichler B.J., Kneilling M., la Fougère C., Forchhammer S., Metzler G., Bauer J., Weide B., Schippert W., Stevanović S, & Löffler M.W. (2019). A new synthetic toll-like receptor 1/2 ligand is an efficient adjuvant for peptide vaccination in a human volunteer. Journal for Immunotherapy of Cancer, 7, 307.

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Flow Cytometric Analysis of Multimer-Specific T Cells

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Multimers were produced by addition of streptavidin-R-phycoerythrin
conjugate (SAPE, ThermoFisher) successively to a monomer, with 30 min of
incubation while rotating overhead between each SAPE addition. Primed
CD8⁺ T cells were stained with Life/Dead Fixable Aqua
(ThermoFisher), CD8-PE-Cy7 (Beckmann Coulter, Brea, CA) and multimers and
measured using a FACSCanto (BD Biosciences, Franklin Lakes, NJ). Unstimulated
and unstained PBMCs and ionomycin/phorbol 12-myristate-13-acetate (PMA) were
used as controls.
Staining was only considered specific when at least a population of 0.5%
of multimer-specific T cells was detectable.

Löffler M.W., Kowalewski D.J., Backert L., Bernhardt J., Adam P., Schuster H., Dengler F., Backes D., Kopp H.G., Beckert S., Wagner S., Königsrainer I., Kohlbacher O., Kanz L., Königsrainer A., Rammensee H.G., Stevanović S, & Haen S.P. (2018). Mapping the HLA-ligandome of Colorectal Cancer Reveals an Imprint of Malignant Cell Transformation. Cancer research, 78(16), 4627-4641.

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Immunophenotyping of CSF Immune Cells

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Routine CSF parameters CSF was drawn as part of routine clinical work-up. Peripheral blood was taken immediately after lumbar puncture. CSF cells were counted manually in a Fuchs Rosenthal chamber. CSF glucose and lactate levels were measured by an enzymatic-amperometric method using chip-sensor technology (EKF Biosen C-Line). Albumin CSF/serum ratios and intrathecal immunoglobulin synthesis were determined by nephelometry (Siemens ProSpec®). Fluorescence-associated cell sorting (FACS) of CSF immune cells was performed as described previously. 18 The following antibodies were used for staining: cluster of differentiation (CD) 45, V450; CD3, APC-Cy7; CD4, PerCP (all BD Bioscience); CD8, PE-Cy7; CD14, FITC; CD19, ECD; CD138, PE; and CD56, APC (all Beckman Coulter). We stained CD4+ T cells (CD45 + CD3 + CD4 +), CD8 + T cells (CD45 + CD3 + CD8+), monocytes (CD45+CD14+), natural killer (NK) cells (CD45+CD56+), B cells (CD45+CD19+CD138-) and plasmablasts (CD45+CD19+CD138+). The percentage of each subpopulation was determined in relation to all CD45 positive single cells using FlowJo (version 10.1). In the discovery cohort, B cells and plasmablasts staining was missing in seven patients and monocytes staining was missing in two patients. CSF parameters are summarized in Table 2.

Biberacher V., Schmidt P., Selter R.C., Pernpeinter V., Kowarik M.C., Knier B., Buck D., Hoshi M.M., Korn T., Berthele A., Kirschke J.S., Zimmer C., Hemmer B, & Mühlau M. (2018). Fatigue in multiple sclerosis: Associations with clinical, MRI and CSF parameters. Multiple sclerosis (Houndmills, Basingstoke, England), 24(8).

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