Lane 1: Wild-type HEK-293T cell lysate 20 ug
Lane 2: NDUFS3 knockout HEK-293T cell lysate 20 ug
Lanes 1 - 2:Merged signal (red and green). Green - ab177471 observed at 27 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab177471 was shown to react with NDUFS3 in wild-type HEK-293T cells in western blot with loss of signal observed in NDUFS3 knockout cell line ab266419 (NDUFS3 knockout cell lysate ab257556). Wild-type and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab177471 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 HEK-293T cell lysate 20 ug
Lane 2: NDUFS3 knockout HEK-293T cell lysate 20 ug
Lanes 1 - 2:Merged signal (red and green). Green - ab183733 observed at 27 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab183733 was shown to react with NDUFS3 in wild-type HEK-293T cells in western blot with loss of signal observed in NDUFS3 knockout cell line ab266419 (NDUFS3 knockout cell lysate ab257556). Wild-type and NDUFS3 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab183733 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4^°C at a 1 in 10000 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 HEK-293T cell lysate (20µg)

Lane 2: NDUFS3 knockout HEK-293T cell lysate (20µg)
Lane 3: HepG2 cell lysate (20µg)
Lane 4: HL60 cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab183733 observed at 30 kDa. Red - loading control ab7291 observed at 50 kDa.
ab183733 Rabbit monoclonal [EPR12781] to NDUFS3 was shown to specifically react with NDUFS3 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266419 (knockout cell lysate ab257556) was used. Wild-type and NDUFS3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab183733 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4^°C at 1 in 10000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Lane 1: Wild-type HEK-293T cell lysate (20µg)

Lane 2: NDUFS3 knockout HEK-293T cell lysate (20µg)
Lane 3: HepG2 cell lysate (20µg)
Lane 4: HL60 cell lysate (20µg)
Lanes 1- 4: Merged signal (red and green). Green - ab177471 observed at 30 kDa. Red - loading control ab7291 observed at 50 kDa.
ab177471 Rabbit monoclonal [EPR12782] to NDUFS3 - C-terminal was shown to specifically react with NDUFS3 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266419 (knockout cell lysate ab257556) was used. Wild-type and NDUFS3 knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab177471 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4^°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

Homozygous: 19 bp deletion in exon 1