Validation Data Gallery

WB result of TNF-alpha antibody (17590-1-AP, 1:2000) with loading-control GAPDH antibody (10494-1-AP, 1:200000) incubated at room temperature for 1.5 hours. Sample 1: THP-1 cells were treated with 80 nM PMA overnight. Sample 2: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours. Sample 3: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (PTG) for the last 3 hours. Sample 4: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (Competitor A) for the last 3 hours.
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WB result of TNF-alpha antibody (17590-1-AP, 1:2000) with loading-control GAPDH antibody (10494-1-AP, 1:200000) incubated at room temperature for 1.5 hours. Sample 1: THP-1 cells were treated with 80 nM PMA overnight. Sample 2: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours. Sample 3: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (PTG) for the last 3 hours. Sample 4: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (Competitor A) for the last 3 hours.
WB result of IL-6 antibody (21865-1-AP, 1:600) with loading-control GAPDH antibody (10494-1-AP, 1:200000) incubated at room temperature for 1.5 hours. Sample 1: THP-1 cells were treated with 80 nM PMA overnight. Sample 2: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours. Sample 3: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (PTG) for the last 3 hours. Sample 4: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (Competitor A) for the last 3 hours.
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WB result of IL-6 antibody (21865-1-AP, 1:600) with loading-control GAPDH antibody (10494-1-AP, 1:200000) incubated at room temperature for 1.5 hours. Sample 1: THP-1 cells were treated with 80 nM PMA overnight. Sample 2: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours. Sample 3: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (PTG) for the last 3 hours. Sample 4: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (Competitor A) for the last 3 hours.
1X10^6 cells were intracellularly stained with 0.4 ug Anti-Human TNF-alpha antibody (17590-1-AP) and Multi-rAb™ CoraLite® Plus 647-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with 0.2% Triton. Sample 1: THP-1 cells were treated with 80 nM PMA overnight. Sample 2: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours. Sample 3: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (PTG) for the last 3 hours. Sample 4: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (Competitor A) for the last 3 hours.
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1X10^6 cells were intracellularly stained with 0.4 ug Anti-Human TNF-alpha antibody (17590-1-AP) and Multi-rAb™ CoraLite® Plus 647-Goat Anti-Rabbit Recombinant Secondary Antibody (H+L), or 0.4 ug Isotype Control. Cells were fixed with 4% PFA and permeabilized with 0.2% Triton. Sample 1: THP-1 cells were treated with 80 nM PMA overnight. Sample 2: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours. Sample 3: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (PTG) for the last 3 hours. Sample 4: THP-1 cells were treated with 80 nM PMA overnight, replaced the culture medium with 100 ng/mL LPS for 6 hours with addition of 2 uL/mL Protein Transport Inhibitor (Competitor A) for the last 3 hours.