Lane 1: Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 2: Wild-type A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lane 3: IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 4: IL-6 knockout A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab233706 observed at 25 kDa, possible non-specific binding at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab233706 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type and IL-6 knockout A549 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab233706 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Lane 1: Wild-type A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 2: Wild-type A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lane 3: IL-6 knockout A549 Brefeldin A (ab120299)-treated (5ug/ml, 4h) cell lysate 30 ug
Lane 4: IL-6 knockout A549 IL-1β (ab259387) (20 ng/ml, 24h) and Brefeldin A (ab120299)-treated (5 ug/ml for the last 4h) cell lysate 30 ug
Lanes 1 - 4: Merged signal (red and green). Green - ab214429 observed at 25 kDa, possible non-specific binding at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab214429 was shown to react with IL-6 in wild-type A549 cells in western blot with loss of signal observed  in IL-6 knockout cell line ab273751 (knockout cell lysate ab275501). Wild-type A549 and IL-6 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab214429 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Allele-1: 97 bp deletion in exon 3.