PBMCs were stimulated with either PMA/Iono (Cell stimulation cocktail, eBioscience), plate-bound CD3/CD49d/soluble CD28 (at 10 μg/mL, 5 μg/mL, and 5 μg/mL respectively) or with peptides or their mixtures [Influenza Matrix peptide57–66 (AnaSpec, San Jose, CA), EBV BZLF-1 peptide pool, CMV pp65 peptide pool (both Miltenyi Biotec, San Diego, CA) or HIV-1 PTE Gag peptide pool43 (link) (obtained from the NIH AIDS reagent program)] for 3 hrs in the presence of brefeldin A (eBioscience). All peptides were used according to manufacturer’s instructions at approximately 1 μg/mL of each peptide; peptide mixtures were constructed as sets of 15-mers overlapped by 9 amino acids. Phenotype and cytokine production were evaluated by flow cytometry as described above. In the phospho ERK blocking experiment, MAPK inhibitor U0126 (Tocris Bioscience, Bristol, UK) was added to PBMCs for 3 h prior stimulation with PMA/Iono.
Mapk inhibitor u0126
Manufactured by Bio-Techne
Sourced in United Kingdom
U0126 is a potent and selective inhibitor of the mitogen-activated protein kinase (MAPK) kinases, MEK1 and MEK2. It inhibits the activation of ERK1/2 by binding to the inactive forms of MEK1/2, preventing their activation by upstream kinases.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using mapk inhibitor u0126
1
Multimodal PBMC Stimulation and Analysis
2
Multimodal PBMC Stimulation and Analysis
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PBMCs were stimulated with either PMA/Iono (Cell stimulation cocktail, eBioscience), plate-bound CD3/CD49d/soluble CD28 (at 10 μg/mL, 5 μg/mL, and 5 μg/mL respectively) or with peptides or their mixtures [Influenza Matrix peptide57–66 (AnaSpec, San Jose, CA), EBV BZLF-1 peptide pool, CMV pp65 peptide pool (both Miltenyi Biotec, San Diego, CA) or HIV-1 PTE Gag peptide pool43 (link) (obtained from the NIH AIDS reagent program)] for 3 hrs in the presence of brefeldin A (eBioscience). All peptides were used according to manufacturer’s instructions at approximately 1 μg/mL of each peptide; peptide mixtures were constructed as sets of 15-mers overlapped by 9 amino acids. Phenotype and cytokine production were evaluated by flow cytometry as described above. In the phospho ERK blocking experiment, MAPK inhibitor U0126 (Tocris Bioscience, Bristol, UK) was added to PBMCs for 3 h prior stimulation with PMA/Iono.
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