Conjugated antibodies and staining reagents included MIP-1β-PE, CD3-PB, CD3-PE-Cy7, CD3-PerCP-Cy5.5, CD3-APC-Cy7, CD3-Horizon V450, CD4-PerCP-Cy5.5, CD4-AmCyan, CD4-FITC, CD8α-APC, CD8α-APC-Cy7, CD8α-AlexaFluor700, CD8α-FITC, CD8α-APC-H7, CD69- ECD (Beckman Coulter), CD20-Horizon V450, CCR7-FITC (R&D Systems), CD95-APC, CCR7-PerCP-Cy5.5, CD28-PE-Cy7 (eBioscience), granzyme B-AlexaFluor700, perforin-FITC (MabTech), IFNγ-PE-Cy7, TNFα- AlexaFluor700, IL-2-APC, CD95-PE, CD95-APC, and Aqua LIVE/DEAD Fixable Dead Cell Stain (Invitrogen). All reagents are from BD Biosciences unless indicated otherwise. For construction of monomers and tetramers, the following peptides were synthesized and purified to >95% by HPLC by New England Peptide LLC: p11C (CTPYDINQM), p54AS (TVPWPNASL), p54E660 (TVPWPNETL), p68A (STPPLVRLV), and TL8 (TTPESANL). The monomers and tetramers were prepared as previously described [85] (link), [86] (link). Tetramers were prepared using either streptavidin-PE (Prozyme), -APC (Prozyme), -AlexaFluor488 (Invitrogen), or -Qdot655 (Invitrogen). Monomers used in surface plasmon resonance studies were further quantified using an RC DC protein kit (Bio-Rad).
Cd28 pe cy7
Manufactured by Thermo Fisher Scientific
Sourced in United States
The CD28-PE-Cy7 is a flow cytometry reagent that can be used to detect the expression of the CD28 surface marker on cells. It consists of an anti-CD28 antibody conjugated with the fluorescent dyes Phycoerythrin (PE) and Cyanine 7 (Cy7).
Automatically generated - may contain errors
Lab products found in correlation
Qdot 655, by Thermo Fisher Scientific (1 mentions)
Ccr7 fitc, by R&D Systems (1 mentions)
Alexa fluor 488, by Thermo Fisher Scientific (1 mentions)
Live dead fixable aqua dead cell stain, by Thermo Fisher Scientific (1 mentions)
Cd69 ecd, by Beckman Coulter (1 mentions)
Cd3 percp cy5, by Beckman Coulter (1 mentions)
Cd4 fitc, by Beckman Coulter (1 mentions)
Cd4 percp cy5, by Beckman Coulter (1 mentions)
Ccr7 percp cy5, by Thermo Fisher Scientific (1 mentions)
Cd3 pe cy7, by Beckman Coulter (1 mentions)
Cd95 apc, by Thermo Fisher Scientific (1 mentions)
Perforin fitc, by Mabtech (1 mentions)
Mip 1β pe, by Beckman Coulter (1 mentions)
Anti cd8 apc, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by EUROIMMUN (1 mentions)
Cd45ro pe, by Thermo Fisher Scientific (1 mentions)
Cd57 fitc, by Thermo Fisher Scientific (1 mentions)
2 protocols using cd28 pe cy7
1
Multiparameter Flow Cytometry Assay
2
Immunophenotyping of NK and CD8+ T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To identify NK and CD8+ T cells subsets, Blood samples (about 10 ml) were
collected in EDTA-containing tubes. The peripheral blood mononuclear cells (PBMCs) were
first isolated using Ficoll-Hypaque (Innotrain, Germany) density gradient centrifugation
(1000×g at room temperature) and then washed two times with phosphate-buffered saline
(PBS, Euroimmun, Germany). Centrifugation of samples was at 300 ×g and 4°C. Subsequently,
isolated PBMCs (106 cells/100 µl) were stained with anti-CD8-APC, CD45RO-PE,
CD27-Percp-eFlour780, CD28-Pe-Cy7, CD57-FITC, and CD56-PerCp-eFlour710 monoclonal
antibodies (mAbs) (all from eBioscience, USA) and incubated at 4°C for 30 minutes. The
samples were fixed with formaldehyde and analyzed within 24 hours by flow cytometer (BD
FACSAria, USA). At least 50,000 events were counted for each sample.
collected in EDTA-containing tubes. The peripheral blood mononuclear cells (PBMCs) were
first isolated using Ficoll-Hypaque (Innotrain, Germany) density gradient centrifugation
(1000×g at room temperature) and then washed two times with phosphate-buffered saline
(PBS, Euroimmun, Germany). Centrifugation of samples was at 300 ×g and 4°C. Subsequently,
isolated PBMCs (106 cells/100 µl) were stained with anti-CD8-APC, CD45RO-PE,
CD27-Percp-eFlour780, CD28-Pe-Cy7, CD57-FITC, and CD56-PerCp-eFlour710 monoclonal
antibodies (mAbs) (all from eBioscience, USA) and incubated at 4°C for 30 minutes. The
samples were fixed with formaldehyde and analyzed within 24 hours by flow cytometer (BD
FACSAria, USA). At least 50,000 events were counted for each sample.
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