Anti-RIP3 antibody [EPR24374-135] - BSA and Azide free

• WB

Lab

Western blot - Anti-RIP3 antibody [EPR24374-135] - BSA and Azide free (AB305055)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab305054).

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='/en-us/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 (<a href='/en-us/products/cell-lines/human-ripk3-knockout-thp-1-cell-line-ab280065'>ab280065</a>) 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

• WB

Supplier Data

Western blot - Anti-RIP3 antibody [EPR24374-135] - BSA and Azide free (AB305055)

This data was developed using ab305054, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : MCF7, HepG2, A549 (PMID : 25675296) Lysates were freshly made and used immediately to minimize protein degradation. Exposure time : 26 seconds

All lanes:

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

Lane 1:

U937 (human histiocytic lymphoma monocyte), whole cell lysate at 10 µg

Lane 2:

THP-1 (human monocytic leukemia monocyte), whole cell lysate at 10 µg

Lane 3:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 10 µg

Lane 4:

HepG2 (human hepatocellar carcinoma epithelial cell), whole cell lysate at 10 µg

Lane 5:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 57 kDa

Observed band size: 56 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-RIP3 antibody [EPR24374-135] - BSA and Azide free (AB305055)

This data was developed using ab305054, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : HeLa (PMID : 26269957, PMID : 28179994) Lysates were freshly made and used immediately to minimize protein degradation. Exposure time : 3 minutes

All lanes:

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

Lane 1:

HT-29 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 57 kDa

Observed band size: 56 kDa

false

Exposure time: 3min