Anti-Rubicon/Baron antibody [EPR26996-20]

• WB

Lab

Western blot - Anti-Rubicon/Baron antibody [EPR26996-20] (AB307720)

Western blot : Anti-Rubicon/Baron antibody [EPR26996-20] ab307720 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 130 kDa in Wild-type A549 cell lysates with no signal observed at this size in RUBCN knockout A549 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 & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-Rubicon/Baron antibody [EPR26996-20] (ab307720) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

RUBCN knockout A549 at 20 µg

Lane 3:

Raji at 20 µg

Lane 4:

HepG2 at 20 µg

Lane 5:

THP-1 at 20 µg

Secondary

Lanes 1 - 5:

Goat anti-Rabbit 800CW at 1/20000 dilution

Lanes 1 - 5:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 109 kDa,130 kDa

Observed band size: 130 kDa,51 kDa

false

• WB

Lab

Western blot - Anti-Rubicon/Baron antibody [EPR26996-20] (AB307720)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : THP-1 (PMID : 22423966). In lane 1-3, the lysates were stored at -80°C prior to Western Blotting. The bands beneath the target band (150 kDa) are likely to be degradation products. In lanes 4-5, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-Rubicon/Baron antibody [EPR26996-20] (ab307720) at 1/1000 dilution

Lane 1:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 2:

K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Raji whole cell lysate at 20 µg

Lane 5:

THP-1 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 1/20000 dilution

Observed band size: 150 kDa

false

Exposure time: 180s