For intracellular cytokine staining, T cells were incubated for 4 h at 37°C in RPMI medium 1640 + 10% FCS in the presence of 10 ng/ml PMA (Sigma-Aldrich, St. Louis, MO), 1 µM ionomycin (EMD Chemicals, Gibbstown, NJ) and with 10 µg/ml brefeldin A (Sigma-Aldrich, St. Louis, MO). Cells were then stained for surface markers as described above. Intracellular staining was preformed using the Cytofix/Cytoperm kit (eBioscience). Surface stained cells were fixed overnight and permeabilized for 45 m, then stained for APC-conjugated anti–IL2 (JESS-5H4), PE-Cy7–conjugated anti–IFNγ (XMG1.2), and eFluor450 anti_TNF (MP6-XT22) (eBiosciences). For in vitro proliferation and ELISA assays, sorted T cells were CFSE-labeled and then stimulated in plates coated (O/N) with CD3 and CD28 antibodies (both eBiosciences) at 1:10 ratio for 8 hours or 96 hours in complete media. The T cells were subsequently analyzed for their mean cell division response and supernatants from each stimulated population (Naive, Treg cell, Teff/mem, anergic) were assayed using ELISA and previously described10 (link).
Pe cy7 conjugated anti ifnγ xmg1.2
Manufactured by Thermo Fisher Scientific
The PE-Cy7–conjugated anti–IFNγ (XMG1.2) is a laboratory reagent used for the detection and analysis of interferon gamma (IFNγ) in flow cytometry applications. The product consists of a monoclonal antibody specific to IFNγ that is conjugated to the PE-Cy7 fluorescent dye.
Automatically generated - may contain errors
Lab products found in correlation
Apc conjugated anti il2 jess 5h4, by Thermo Fisher Scientific (2 mentions)
Efluor450 anti tnf mp6 xt22, by Thermo Fisher Scientific (2 mentions)
Cd28 antibody, by Thermo Fisher Scientific (2 mentions)
Cytofix cytoperm kit, by Thermo Fisher Scientific (2 mentions)
Brefeldin a, by Merck Group (2 mentions)
Phorbol myristate acetate (pma), by Merck Group (2 mentions)
2 protocols using pe cy7 conjugated anti ifnγ xmg1.2
1
Intracellular Cytokine Staining of T Cells
2
Intracellular Cytokine Staining of T Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For intracellular cytokine staining, T cells were incubated for 4 h at 37°C in RPMI medium 1640 + 10% FCS in the presence of 10 ng/ml PMA (Sigma-Aldrich, St. Louis, MO), 1 µM ionomycin (EMD Chemicals, Gibbstown, NJ) and with 10 µg/ml brefeldin A (Sigma-Aldrich, St. Louis, MO). Cells were then stained for surface markers as described above. Intracellular staining was preformed using the Cytofix/Cytoperm kit (eBioscience). Surface stained cells were fixed overnight and permeabilized for 45 m, then stained for APC-conjugated anti–IL2 (JESS-5H4), PE-Cy7–conjugated anti–IFNγ (XMG1.2), and eFluor450 anti_TNF (MP6-XT22) (eBiosciences). For in vitro proliferation and ELISA assays, sorted T cells were CFSE-labeled and then stimulated in plates coated (O/N) with CD3 and CD28 antibodies (both eBiosciences) at 1:10 ratio for 8 hours or 96 hours in complete media. The T cells were subsequently analyzed for their mean cell division response and supernatants from each stimulated population (Naive, Treg cell, Teff/mem, anergic) were assayed using ELISA and previously described10 (link).
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!