Human RIPK3 knockout THP-1 cell line

• WB

Lab

Western blot - Human RIPK3 knockout THP-1 cell line (AB280065)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab325636 was shown to bind specifically to RIPK3. Target of interest was observed at 37-50 kDa in wild-type THP-1 cell lysates (lane 1) with no signal observed at this size in RIPK3 knockout cell line (lane 2)(knockout cell line ab280065).

In Western blot, Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (ab176842) (1 : 100000) (17KDa).

All lanes:

Western blot - Anti-RIP3 antibody [RM1355] (<a href='/ko/products/primary-antibodies/rip3-antibody-rm1355-ab325636'>ab325636</a>) at 1/1000 dilution

Lane 1:

Wilde-type THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 2:

Western blot - Human RIPK3 knockout THP-1 cell line (ab280065) at 20 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

Observed band size: 37-56 kDa,17 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Human RIPK3 knockout THP-1 cell line (AB280065)

Western blot : Anti-RIP3 antibody [EPR24374-135] ab305054 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 57 kDa in Wild-type THP-1 cell lysates with no signal observed at this size in RIPK3 knockout THP-1 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-RIP3 antibody [EPR24374-135] (<a href='/ko/products/primary-antibodies/rip3-antibody-epr24374-135-ab305054'>ab305054</a>) at 1/1000 dilution

Lane 1:

Wild-type THP-1 whole cell lysate at 20 µg

Lane 2:

Western blot - Human RIPK3 knockout THP-1 cell line (ab280065) at 20 µg

Lane 3:

Jurkat whole cell lysate at 20 µg

Lane 4:

HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 57 kDa

Observed band size: 57 kDa

false