Anti-c-Rel antibody [EPR25178-58] - BSA and Azide free

• WB

Supplier Data

Western blot - Anti-c-Rel antibody [EPR25178-58] - BSA and Azide free (AB309132)

This data was developed using ab309131, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : ½ volume of Odyssey Blocking Buffer (TBS)+ ½ volume of TBS Lysates at 20 µg per lane. The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-c-Rel antibody (ab309131) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab309131 was shown to bind specifically to c-Rel. Target of interest was observed at 78 kDa, in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in c-Rel knockout cell line (lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged. Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-c-Rel antibody [EPR25178-58] (<a href='/en-us/products/primary-antibodies/c-rel-antibody-epr25178-58-ab309131'>ab309131</a>) at 1/1000 dilution

Lane 1:

293T cells transfected with a human LTBR expression vector containi a His-tag whole cell lysate

Lane 2:

c-Rel knockout HAP1 cell lysate

Lane 3:

Raji (human burkitt's lymphoma b lymphocyte) whole cell lysate

Lane 4:

HuT-78 (human sezary syndrome cutaneous t lymphocyte) whole cell lysate

Secondary

All lanes:

Goat Anti-Rabbit IgG H&L (800CW) and Goat Anti-Mouse IgG H&L (680RD) at 1/10000 dilution

Observed band size: 78 kDa

false

• WB

Supplier Data

Western blot - Anti-c-Rel antibody [EPR25178-58] - BSA and Azide free (AB309132)

This data was developed using ab309131, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Exposure time : Lanes 1-3 : 26 seconds; Lanes 4-7 : 10 seconds.

All lanes:

Western blot - Anti-c-Rel antibody [EPR25178-58] (<a href='/en-us/products/primary-antibodies/c-rel-antibody-epr25178-58-ab309131'>ab309131</a>) at 1/1000 dilution

Lane 1:

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

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

Rat thymus tissue lysate at 20 µg

Lane 5:

Mouse thymus tissue lysate at 20 µg

Lane 6:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 7:

Raji (human burkitts lymphoma b lymphocyte) 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: 78 kDa,68 kDa

false