Tumor necrosis factor-α (TNF-α), interleukin (IL)-17A, IL-10, IL-6 enzyme-linked immunosorbent assay (ELISA) kits, fluorescent-labeled monoclonal antibodies anti-CD4-fluorescein isothiocyanate (FITC), anti-IgG1-FITC, anti-CD25-Phycoerythrin (PE), anti-IgG1-PE, anti-FoxP3-Allophycocyanin (APC), anti-Interferon (IFN)-gamma-PE, anti-IgG1-APC, and anti-IL-17-APC were bought from eBiosciences (San Jose, CA, USA). FIX & PERM medium was obtained from Invitrogen (California, USA). Leukocyte Activation Cocktail was obtained from BD (New York, USA), Catalog No.550583, containing phorbol 12-myristate 13-acetate (PMA), a calcium ionophore (ionomycin), and the protein transport inhibitor BD GolgiPlug™ (Brefeldin A, New York, USA).
Anti cd4 fluorescein isothiocyanate fitc
Manufactured by Thermo Fisher Scientific
Sourced in United States
Anti-CD4-fluorescein isothiocyanate (FITC) is a fluorescently labeled antibody that binds to the CD4 protein on the surface of T cells. It is used in flow cytometry applications to identify and quantify CD4+ T cells in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Golgiplug, by BD (1 mentions)
Anti foxp3 allophycocyanin apc, by Thermo Fisher Scientific (1 mentions)
Leukocyte activation cocktail, by BD (1 mentions)
Anti il 17 apc, by Thermo Fisher Scientific (1 mentions)
Anti cd25 phycoerythrin pe, by Thermo Fisher Scientific (1 mentions)
Anti igg1 pe, by Thermo Fisher Scientific (1 mentions)
Fix perm medium, by Thermo Fisher Scientific (1 mentions)
Anti cd4 fitc, by Thermo Fisher Scientific (1 mentions)
Anti foxp3 pe, by Thermo Fisher Scientific (1 mentions)
Anti cd25 apc, by Thermo Fisher Scientific (1 mentions)
Anti il 17a apc, by Thermo Fisher Scientific (1 mentions)
Cell stimulation cocktail, by Thermo Fisher Scientific (1 mentions)
Aria 2, by BD (1 mentions)
Anti cd3 pe, by Thermo Fisher Scientific (1 mentions)
Anti cd38 pe cy5, by BioLegend (1 mentions)
Accuri c6 flow cytometer, by BD (1 mentions)
Anti cd19 pe, by Beckman Coulter (1 mentions)
Fixation permeabilization buffer solution, by Thermo Fisher Scientific (1 mentions)
Anti cd25 pe cy5, by Thermo Fisher Scientific (1 mentions)
3 protocols using anti cd4 fluorescein isothiocyanate fitc
1
Cytokine and Lymphocyte Profiling Protocol
2
Detecting Treg and Th17 Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For detection of Treg cells, PBMCs were stained with surface markers anti-CD4-fluorescein isothiocyanate (FITC; 11-0049-80, eBioscience) and anti-CD25-APC (17-0259-42, eBioscience). After fixation and permeabilization, intracellular FOXP3 staining was conducted using anti-FOXP3-PE (12-4776-42, eBioscience). For detection of Th17 cells, PBMCs were stimulated with cell stimulation cocktail (plus protein transport inhibitors; 00-4975-93, Invitrogen) for 4 h. Cells were then harvested and rinsed with PBS. Cell viability was detected by FVS780 staining (565388, BD Biosciences, San Jose, CA, USA), followed by surface staining with anti-CD3-PE (12-0038-42, eBioscience) and anti-CD4-FITC (11-0049-80, eBioscience). Subsequently, cells were stained with anti-IL17A-APC (17-7179-42, eBioscience) following fixation and permeabilization. FACS was performed using BD Aria II (BD Biosciences), and data were analyzed using FlowJo software (Tree Star, Ashland, OR, USA).
3
Phenotyping of Regulatory T and B Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Approximately 1x10 6 PBMC were deposited into flow cytometry tubes (Falcon, Becton Dickinson, Oxnard, California). To identify Treg cells, surface labeling was performed with anti-CD4-fluorescein isothiocyanate (FITC) (clone: OKT4, eBioscience, San Diego, CA) and anti-CD25-PECy5 (clone: BC96 eBioscience) antibodies. The cells were fixed with 2% paraformaldehyde, permeabilized with the fixation/permeabilization buffer solution (eBioscience, San Diego, CA) according to the manufacturer's recommendations, and subsequently incubated with anti-FOXP3-PE antibody (clone: 259D Biolegend). Finally, the cells were washed and resuspended in phosphate-buffered saline (PBS) and read on an Accuri C6 flow cytometer (BD Biosciences). For the phenotyping of the Breg cells, anti-CD19-PE (Beckman Coulter, Brea, CA, USA), anti-CD24-Alexa-fluor-647 (clone: ML5 BioLegend) and anti-CD38-PECy5 (clone: HIT2 BioLegend) antibodies were added for 30 minutes at room temperature.
Subsequently, the cells were washed with PBS and fixed with 2% paraformaldehyde. The samples were read in a flow cytometer.
Subsequently, the cells were washed with PBS and fixed with 2% paraformaldehyde. The samples were read in a flow cytometer.
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!