Anti-TRF1 antibody [EPR28580-71]

• WB

Lab

Western blot - Anti-TRF1 antibody [EPR28580-71] (AB323380)

Western blot : Anti-TRF1 antibody [EPR28580-71] ab323380 staining at 1/1000 dilution, shown in green; Mouse anti-CANX ab238078 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 55 kDa in Wild-type A549 cell lysates with no signal observed at this size in TERF1 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 5pc 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-TRF1 antibody [EPR28580-71] (ab323380) at 1/1000 dilution

Lane 1:

Wild-type A549 whole cell lysate at 20 µg

Lane 2:

TERF1 knockout A549 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Lane 4:

HEK-293 whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 50 kDa

Observed band size: 55 kDa

false

• WB

Supplier Data

Western blot - Anti-TRF1 antibody [EPR28580-71] (AB323380)

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

This antibody was tested on frozen and fresh HeLa and Jurkat whole cell lysate and it was concluded that TRF1 is the easy target for degradation.

The bands beneath the target band are likely to be degraded target fragments.

To minimize protein degradation, cells (lanes 7-8) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-6 : 48 seconds; Lanes 7-8 : 92 seconds

All lanes:

Western blot - Anti-TRF1 antibody [EPR28580-71] (ab323380) at 1/1000 dilution

Lanes 1 and 7:

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

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lanes 3 and 8:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 4:

LN-229 (human brain glioblastoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 6:

K-562 (human chronic myelogenous leukemia lymphoblast) 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: 60 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Anti-TRF1 antibody [EPR28580-71] (AB323380)

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

The bands beneath the target band are likely to be degraded target fragments.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-TRF1 antibody [EPR28580-71] (ab323380) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting TRF1 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: 60 kDa,36 kDa

false

Exposure time: 48s