Anti-RIP 抗体 [EPR4689-100]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-RIP antibody [EPR4689-100] (AB178420)

Intracellular flow cytometric analysis of permeabilized HeLa cells labeling RIP with ab178420 at 1/10 dilution (red) compared with a rabbit IgG negative control (green).

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

Western blot : Anti-RIPK1 antibody [EPR4689-100] (ab178420) 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, ab178420 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 [EPR4689-100] (ab178420) 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

• WB

Unknown

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

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 to 4 : Merged signal (red and green). Green - ab178420 observed at 78 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab178420 was shown to react with RIP in wild-type HAP1 cells along with additional cross-reactive bands. No band was observed when RIP knockout samples were examined. Wild-type and RIP knockout samples were subjected to SDS-PAGE. ab178420 and ab8245 (loading control to GAPDH) were both diluted 1/1000 and 1/10,000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) ab216776 secondary antibodies at 1/10,000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420)

Predicted band size: 75 kDa

false

• WB

Lab

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution

Lanes 1 - 4:

Western blot - Human RIPK1 knockout THP-1 cell line (<a href='/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

Supplier Data

Western blot - Anti-RIP antibody [EPR4689-100] (AB178420)

All lanes:

Western blot - Anti-RIP antibody [EPR4689-100] (ab178420) at 1/1000 dilution

Lane 1:

Raji cell lysate at 10 µg

Lane 2:

HeLa cell lysate with Staurosporine at 10 µg

Lane 3:

HeLa cell lysate at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 75 kDa

false