After 2 days of culture, tonsil cells were stimulated with 50 ng/mL of phorbol 12-myristate 13-acetate (PMA, Sigma P8139) and 1 μg/mL of ionomycin (Sigma I3909) in the presence of protein transport inhibitor containing monensin (BD GolgiStop) for 5 hours. Cryopreserved rhesus macaque cells were rested for 24 hours (viability 40-80%) in R10 and then stimulated in the same manner for 5 hours. Cells were then harvested, blocked and stained for surface markers as above, and then fixed and permeabilized using BD CytoFix/Cytoperm kit (554714) according to manufacturers’ instructions. Cells were then stained at 4 °C for 30 minutes with IL-4-PercpCy5.5-8D4-8 (BD 561234), IL-10-eFluor450-JES3-9D7 (eBioscience 48-7108), IL-21-AF647-3A3-N2.1 (BD 560493), and TGFβ-1-PE(LAP)-27232 (R&D FAB2463P). All antibodies were used at one test per 106 cells. All cytokine analyses were normalized to a mock-spinoculated control that received monensin but was unstimulated.
Il 21 af647 3a3 n2
Manufactured by BD
IL-21-AF647-3A3-N2.1 is a fluorescently labeled antibody used for the detection and analysis of Interleukin-21 (IL-21) in biological samples. The antibody is conjugated with the Alexa Fluor 647 dye, providing a bright and photostable fluorescent signal. It can be used in various immunoassay techniques, such as flow cytometry, to quantify IL-21 levels in cell culture, tissue, or other relevant samples.
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Lab products found in correlation
3 protocols using il 21 af647 3a3 n2
1
Rhesus Macaque Tonsil Cell Cytokine Profiling
2
Rhesus Macaque Tonsil Cell Cytokine Analysis
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After 2 days of culture, tonsil cells were stimulated with 50 ng ml−1 of PMA (Sigma, P8139) and 1 μg ml−1 of ionomycin (Sigma, I3909) in the presence of protein transport inhibitor containing monensin (BD GolgiStop) for 5 h. Cryopreserved rhesus macaque cells were rested for 24 h (viability 40–80%) in R10 and then stimulated in the same manner for 5 h. Cells were then harvested, blocked and stained for surface markers as above, and then fixed and permeabilized using BD CytoFix/Cytoperm kit (554714) according to the manufacturer's instructions. Cells were then stained at 4 °C for 30 min with IL-4-PercpCy5.5-8D4–8 (BD, 561234), IL-10-eFluor450-JES3–9D7 (eBioscience, 48–7108), IL-21-AF647-3A3-N2.1 (BD, 560493) and TGF-β-1-PE(LAP)-27232 (R&D, FAB2463P). All antibodies were used at one test per 106 cells. All cytokine analyses were normalized to a mock-spinoculated control that received monensin but was unstimulated.
3
Tonsil Cell Cytokine Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After 2 days of culture, tonsil cells were stimulated with 50 ng/mL of phorbol 12-myristate 13-acetate (PMA, Sigma P8139) and 1 μg/mL of ionomycin (Sigma I3909) in the presence of protein transport inhibitor containing monensin (BD GolgiStop) for 5 hours. Cells were then harvested, blocked and stained for surface markers as above, and then fixed and permeabilized using BD CytoFix/Cytoperm kit (554714) according to manufacturers’ instructions. Cells were then stained at 4°C for 30 minutes and analyzed for IL-2-PE-MQ1-17H12 (BD 559334), IL-10-eFluor450-JES3-9D7 (eBioscience), IL-21-AF647-3A3-N2.1 (BD 560493), and perforin-FITC-Pf344 (Mabtech 3465–7). All cytokine analyses were normalized to a mock-spinoculated control that received monensin but was not stimulated.
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