The following antibodies were used for flow cytometry: FITC anti-human CD3, CD16, and CD8a; PE anti-human CD4, CD8a, CD141, CD14, and CCR7; APC-eFluor780 anti-human HLA-DR; PE-Cy7 anti-human CD11c; PerCP-eFluor710 anti-human CD1c; APC anti-human CD303α; PE-Cy7 anti-human CD4; APC anti-human CD45RA and CFSE (eBioscience, USA); FITC anti-human HLA-A2 and HLA-DR; PE anti-human CD123, CD40, CD80, CD83, and CD86; APC anti-human CD137 (4-1BB); APC anti-human HLA-ABC (BD Bioscience, USA); PE anti-human EGFR; and APC anti-human CD11c (BioLegend, USA).
Anti human cd4 pe cy7
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Anti-human CD4-PE-Cy7 is a fluorochrome-conjugated monoclonal antibody that binds to the CD4 surface antigen expressed on a subset of T lymphocytes. It can be used in flow cytometric analysis to identify and enumerate CD4-positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Pe anti human cd123, by BD (1 mentions)
Apc anti human cd11c, by BioLegend (1 mentions)
Fitc anti human hla a2, by BD (1 mentions)
Anti human cd4 pe, by Thermo Fisher Scientific (1 mentions)
Facs caliber flow cytometer, by BD (1 mentions)
Pe anti human cd8, by BioLegend (1 mentions)
Filcon, by BD (1 mentions)
Tryple, by Thermo Fisher Scientific (1 mentions)
Anti human cd19 apc, by Thermo Fisher Scientific (1 mentions)
Anti human cd11c apc, by Thermo Fisher Scientific (1 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Facs navios, by Beckman Coulter (1 mentions)
Anti human cd127 fitc, by Thermo Fisher Scientific (1 mentions)
Anti human cd25 apc, by Thermo Fisher Scientific (1 mentions)
Anti mouse cd4 pe cy7, by Thermo Fisher Scientific (1 mentions)
Facsverse flow cytometer, by BD (1 mentions)
Anti mouse ifnγ apc, by BioLegend (1 mentions)
Anti human ifn γ apc, by Thermo Fisher Scientific (1 mentions)
Pe anti mouse cd3, by BioLegend (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
7 protocols using anti human cd4 pe cy7
1
Comprehensive Immune Profiling by Flow Cytometry
2
CD4+ IL9+ T Cell Analysis by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Flow cytometry was used for analyzing the CD4+ IL9+ T subset. Cells were stained with PE-Cy7 anti-human CD4 (eBioscience, San Diego, CA, USA) in a dark place at room temperature for 20 min. After surface staining, cells were fixed, permeabilized and then stained with phycoerythrin (PE) anti-human IL9 (eBioscience, San Diego, CA, USA). To determine cut-offs, matching isotype control antibodies were processed in a similar way. In this study, CD4+ IL9+ T cells were referred to as the Th9 cells. Cells were calculated as a percentage of CD4+ T cells. Stained cells were acquired by FACS-Caliber flow cytometer (BD Bioscience, San Jose, CA, USA) and data were analyzed using FlowJo software 7.6.1 (Tree Star Inc., Ashland, OR, USA). The gating strategy is demonstrated in Figure 1 .
3
Generation of Human T Cells from iPSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A total of 30,000 cells per well of a 12-well plate were cultured on mitomycin C-treated OP9-hDLL1 in stromal culture medium with human cytokines as follow: 20 ng/mL FLT3L, 100 ng/mL SCF, 25 ng/mL IL-2, and 5 ng/mL IL-7 during the first 2 weeks and 10 ng/mL FLT3L, 25 ng/mL IL-2, and 5 ng/mL of IL-7 for the last 2 weeks. The cultures were transferred to fresh OP9-hDLL1 cells every 5 days after the first 10 days. For passaging, the cultures were treated with TrypLe (Thermo Fisher Scientific) and filtered through a 50-μm sterile Filcon (BD). Cells were analyzed after 1 month of culture using hCD45-PeCy5.5 (eBioscience), anti-human CD8-PE (BioLegend), and anti-human CD4-PeCy7 (eBioscience).
4
Lymphocyte Isolation and Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protocol described here was approved by the Institutional Review Board of Hanyang University. Human blood samples were obtained from healthy donors and blood lymphocytes were isolated by density centrifugation using Ficoll-Paque PLUS (GE Healthcare). The isolated lymphocytes were seeded 1.0 × 106 cells per well and the delivery efficiencies of CPP-proteins were analysed. The cells were further stained with 1/800 diluted anti-human CD4-PE-Cy7 (#25-0049), anti-human CD19-APC (#17-0199), anti-human CD11b-PE-Cy7 (#25-0118) or anti-human CD11c-APC (#17-0116) FACS antibodies purchased from eBioscience.
5
Quantifying Regulatory T Cells by Flow Cytometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
At each study visit a blood sample was taken for Treg quantification. Tregs were isolated and quantified at the Pediatric Hematology Oncology Lab, Fondazione IRCCS Policlinico S. Matteo, Pavia. For surface staining PBMCs, collected after overnight incubation in RPMI medium, were first incubated with anti-human CD4-Pe-Cy7, anti-human CD25-APC, anti-human CD127-FITC (eBioscience TM, CA). After cell fixation and membrane permeabilization, intracellular staining with anti-Foxp3-PE (Anti-human Foxp3 staining set PE kit; eBioscienceTM, Carlsbad, CA) was performed. Tregs were measured by CD4 T cells gating on the lymphocyte region (forward/sideward scatter), and the frequency of CD25 + CD127neg Foxp3 + cells was calculated as a proportion of CD4 + T cells on an 8-color flow cytometer (FACS Navios, Beckman Coulter).
6
Multicolor Flow Cytometry Analysis of Immune Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Freshly isolated hepatic lymphocytes separated from liver tissue sections or human PBMCs were stained with various antibodies in the dark at 4°C for 30 min, then washed twice with PBS. After fixation and permeabilization with a Cytofix/Cytoperm kit (BD Biosciences, San Jose, CA, USA) and staining with various antibodies, including anti-mouse CD3-PE (100206, BioLegend), anti-mouse NK1.1-FITC (108706, BioLegend), anti-mouse CD8a-FITC (2002714, Invitrogen), anti-mouse CD4-PE-Cy7 (25-0042-82, eBioscience), anti-mouse IFN-γ-APC (505810, BioLegend), anti-human CD4-PE-Cy7 (25-0049-42, eBioscience), anti-human CD8a-FITC (2518338, Invitrogen), anti-human IFN-γ-APC (17-7319-82, eBioscience). The cells were analyzed by flow cytometry on a BD FACSVerseTM Flow Cytometer (BD Bioscience) and the analysis was used by The Guava Soft software.
7
Comprehensive Immune Cell Analysis in Murine Transplantation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The spleen and bone marrow of the model mice and healthy recipient control mice
were harvested at the 6th or 16th week after BMT. Single cell suspensions from
spleens and bone marrows were labeled with anti-CD3 PE, -CD4 FITC, -CD8 AF700
(BD Pharmingen, San Diego, CA, USA) for T cells, anti-CD19 FITC (BD) for B
cells, anti-CXCR3 BV421 and anti-CD80 FITC for immune molecular examination,
anti-CD4 FITC (BD) and anti-Foxp3 PE (eBioscience, San Diego, CA, USA) for
regulatory T cells (Treg), anti-IFN-γ PE and -IL-17A PE-cy7 (BD) for Th1/Th17
subsets, respectively. Before detecting the secretion of IFN-γ and IL-17A in T
cells, cell suspensions were activated with anti-mouse CD3/CD28 microbeads for 3
hours. The spleen mononuclear cells of mice transplanted with A20 lymphoma cells
were stained with anti-B220 PE and -H-2Kb BV421 (eBioscience) to
examine the ratio of tumor cells. Activated human PBMCs with anti- human
CD3/CD28 beads were stained with anti-human CD3 FITC and -CD69 PE for T cell
activation analysis or with anti-human CD4 PE-cy7, -T-bet PE, -Foxp3 AF647,
-GATA3 APC and -ROR?t PE (eBioscience) for T cell differentiation analysis after
incubation with SHR0302 (1 μM) for 24 h. Fixable viability dye or DAPI (BD) were
used to distinguish live cells from dead cells.
were harvested at the 6th or 16th week after BMT. Single cell suspensions from
spleens and bone marrows were labeled with anti-CD3 PE, -CD4 FITC, -CD8 AF700
(BD Pharmingen, San Diego, CA, USA) for T cells, anti-CD19 FITC (BD) for B
cells, anti-CXCR3 BV421 and anti-CD80 FITC for immune molecular examination,
anti-CD4 FITC (BD) and anti-Foxp3 PE (eBioscience, San Diego, CA, USA) for
regulatory T cells (Treg), anti-IFN-γ PE and -IL-17A PE-cy7 (BD) for Th1/Th17
subsets, respectively. Before detecting the secretion of IFN-γ and IL-17A in T
cells, cell suspensions were activated with anti-mouse CD3/CD28 microbeads for 3
hours. The spleen mononuclear cells of mice transplanted with A20 lymphoma cells
were stained with anti-B220 PE and -H-2Kb BV421 (eBioscience) to
examine the ratio of tumor cells. Activated human PBMCs with anti- human
CD3/CD28 beads were stained with anti-human CD3 FITC and -CD69 PE for T cell
activation analysis or with anti-human CD4 PE-cy7, -T-bet PE, -Foxp3 AF647,
-GATA3 APC and -ROR?t PE (eBioscience) for T cell differentiation analysis after
incubation with SHR0302 (1 μM) for 24 h. Fixable viability dye or DAPI (BD) were
used to distinguish live cells from dead cells.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!