Anti-RIP antibody [EPR4689] - BSA and Azide free

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689] - BSA and Azide free (AB234924)

This data was developed using ab125072, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/ko/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/5000 dilution

All lanes:

Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689] - BSA and Azide free (AB234924)

This data was developed using ab125072, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 cell lysate (20 μg)

Lane 2 : RIP knockout HAP1 cell lysate (20 μg)

Lane 3 : HeLa cell lysate (20 μg)

Lane 4 : Raji cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab125072 (unpurified) observed at 78 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab125072 was shown to specifically react with RIP in wild-type HAP1 cells. No band was observed when RIP knockout samples were examined. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab125072 at a dilution of 1/1000 and ab8245 (loading control to GAPDH) at a dilution of 1/10,000 were incubated overnight at 4°C. 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 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/ko/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>)

Predicted band size: 75 kDa

false

• WB

Unknown

Western blot - Anti-RIP antibody [EPR4689] - BSA and Azide free (AB234924)

This data was developed using ab125072, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/ko/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/1000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Predicted band size: 75 kDa

false

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689] - BSA and Azide free (AB234924)

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/ko/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Human RIPK1 knockout THP-1 cell line (<a href='/ko/products/cell-lines/human-ripk1-knockout-thp-1-cell-line-ab276121'>ab276121</a>)

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

Raji cell lysate at 20 µg

Predicted band size: 75 kDa

Observed band size: 76 kDa

true

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689] - BSA and Azide free (AB234924)

This data was developed using ab125072, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/ko/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/1000 dilution

All lanes:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

false

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689] - BSA and Azide free (AB234924)

This data was developed using the same antibody clone in a different buffer formulation (ab125072).

Western blot : Anti-RIPK1 antibody [EPR4689] (ab125072) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab125072 was shown to bind specifically to RIPK1. A band was observed at 75 kDa in wild-type THP-1 cell lysates with no signal observed at this size in RIPK1 knockout cell line. To generate this image, wild-type and RIPK1 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-RIP antibody [EPR4689] (<a href='/ko/products/primary-antibodies/rip-antibody-epr4689-ab125072'>ab125072</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 cell lysate at 20 µg

Lane 2:

RIPK1 knockout THP-1 cell lysate at 20 µg

Lane 3:

Raji cell lysate at 20 µg

Lane 4:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Predicted band size: 75 kDa

Observed band size: 75 kDa

false