All lanes : Anti-PDGFR beta antibody [42G12] (ab69506) at 1/1000 dilution

Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : PDGFRB knockout SH-SY5Y cell lysate
Lane 3 : Human Skeletal Muscle tissue lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 30 µg per lane.

Performed under reducing conditions.

Observed band size: 170 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab69506 observed at 170 kDa. Red - loading control ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) observed at 55 kDa.

ab69506 was shown to react with PDGFR beta in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRB knockout cell line ab273749 (knockout cell lysate ab275523). Wild-type SH-SY5Y and PDGFRB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab69506 and ab52866 (Rabbit anti-alpha Tubulin antibody [EP1332Y]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye^® 680RD) preabsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes : Anti-PDGFR beta antibody [Y92] - C-terminal (ab32570) at 1/5000 dilution

Lane 1 : Wild-type SH-SY5Y cell lysate
Lane 2 : PDGFRB knockout SH-SY5Y cell lysate
Lane 3 : Human Skeletal Muscle tissue lysate
Lane 4 : HeLa cell lysate

Lysates/proteins at 30 µg per lane.

Performed under reducing conditions.

Observed band size: 170 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab32570 observed at 170 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

ab32570 was shown to react with PDGFRB in wild-type SH-SY5Y cells in Western blot with loss of signal observed in PDGFRB knockout cell line ab273749 (knockout cell lysate ab275523). Wild-type SH-SY5Y and PDGFRB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab32570 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 5000 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 h at room temperature before imaging.

Allele-1: 5 bp deletion in exon 3