Application Notes: Capture antibody and detection antibody by WB KO validation (Detect TNF-alpha KO THP-1 cells)
Human TNFalpha(Tumor Necrosis Factor Alpha) ELISA Kit
orb561883
Lane 1 and 3: Wild THP-1 cells were treated with 80nM TPA, cultured overnight, stimulated with 200ng/ml LPS for 6 hours, then cultured with 300ng/ml Brefeldin A (BFA) for 3 hours, and the cell lysate was collected, and the total protein concentration of 1.55mg/ml was detected by BCA, with 10ug of total protein loading. Lane 2 and 4: KO THP-1 cells were treated with 80nM TPA, cultured overnight, stimulated with 200ng/ml LPS for 6 hours. then, then cultured with 300ng/ml Brefeldin A (BFA) for 3 hours, and the cell lysate was collected, and the total protein concentration of 1.55mg/ml was detected by BCA, with 10ug of total protein loading. Primary antibody: Lane 1 and 2:Capture mouse monoclonal antibody 1ug/ml. Lane 3 and 4:Detection mouse monoclonal antibody 1ug/ml. Secondary antibody: All lanes : HRP-Goat Anti-mouse IgG (H&L) (Catalogue No.:FNSA-0003) at 1/5000 dilution. Molecular weight: 26 kDa.
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