All lanes : Anti-Synaptophysin antibody [EP1098Y] (ab52636) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : Syp knockout HEK293T cell lysate
Lane 3 : Human brain tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 33 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab52636 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.

ab52636 Anti-Synaptophysin antibody [EP1098Y] was shown to specifically react with Synaptophysin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267272 (knockout cell lysate ab257060) was used. Wild-type and Synaptophysin knockout samples were subjected to SDS-PAGE. ab52636 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 10000 dilution for 1 hour at room temperature before imaging.

 

All lanes : Anti-Synaptophysin antibody [YE269] (ab32127) at 1/1000 dilution

Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : Syp knockout HEK293T cell lysate
Lane 3 : Human brain tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 33 kDa
Observed band size: 38 kDa why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab32127 observed at 38 kDa. Red - loading control, ab7291 observed at 50 kDa.

ab32127 Anti-Synaptophysin antibody [YE269] was shown to specifically react with Synaptophysin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267272 (knockout cell lysate ab257060) was used. Wild-type and Synaptophysin knockout samples were subjected to SDS-PAGE. ab32127 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 10000 dilution for 1 hour at room temperature before imaging.

 

Homozygous: 8 bp deletion in exon2

Representative images of SYP knockout HEK293T cells, low and high confluency examples (top left and right respectively) and wild-type HEK293T cells, low and high confluency (bottom left and right respectively) showing typical adherent, epithelial-like morphology. Images were captured at 10X magnification using a EVOS XL Core microscope.