Anti-FADD antibody [OTI1C11]

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-FADD antibody [OTI1C11] (AB119059)

HEK293T cells transfected with either a FADD overexpress plasmid (Red) or empty vector control plasmid (Blue) were immunostained using ab119059 at 1/100 dilution and then analyzed by Flow Cytometry (Intracellular).

• WB

Lab

Western blot - Anti-FADD antibody [OTI1C11] (AB119059)

Lanes 1 - 4 : Merged signal (red and green). Green - ab119059 observed at 25 kDa. Red - loading control, ab181602, observed at 37 kDa.
ab119059 was shown to specifically react with FADD when FADD knockout samples were used. Wild-type and FADD knockout samples were subjected to SDS-PAGE. ab119059 and ab181602 (loading control to GAPDH) were both diluted 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed ab216772 and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-FADD antibody [OTI1C11] (ab119059) at 1/2000 dilution

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

FADD knockout HAP1 cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Predicted band size: 23 kDa

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• WB

Lab

Western blot - Anti-FADD antibody [OTI1C11] (AB119059)

Lanes 1-4 : Merged signal (red and green). Green - ab119059 observed at 25 kDa. Red - loading control ab181602 observed at 37 kDa.

ab119059 Anti-FADD antibody [OTI1C11] was shown to specifically react with FADD in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261817 (knockout cell lysate ab257261) was used. Wild-type and FADD knockout samples were subjected to SDS-PAGE. ab119059 and Anti-GAPDH antibody[EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 in 2000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^® 800CW) preadsorbed (ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^® 680RD) preadsorbed (ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-FADD antibody [OTI1C11] (ab119059) at 1/2000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

FADD knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human FADD knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-fadd-knockout-hela-cell-line-ab261817'>ab261817</a>)

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-h-l-irdye-800cw-preadsorbed-ab216772'>ab216772</a>) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 25 kDa

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• WB

Supplier Data

Western blot - Anti-FADD antibody [OTI1C11] (AB119059)

All lanes:

Western blot - Anti-FADD antibody [OTI1C11] (ab119059) at 1/200 dilution

Lane 1:

HepG2 cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

A549 cell lysate at 10 µg

Lane 4:

Jurkat cell lysate at 10 µg

Lane 5:

MCF7 cell lysate at 10 µg

Predicted band size: 23 kDa

false

• WB

Unknown

Western blot - Anti-FADD antibody [OTI1C11] (AB119059)

All lanes:

Western blot - Anti-FADD antibody [OTI1C11] (ab119059) at 1/2000 dilution

Lane 1:

HEK293T cell lysate transfected with pCMV6-ENTRY control at 5 µg

Lane 2:

HEK293T cell lysate transfected with pCMV6-ENTRY FADD cDNA at 5 µg

Predicted band size: 23 kDa

false