Anti-c-Rel antibody [EPR2559(2)] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-c-Rel antibody [EPR2559(2)] - BSA and Azide free (AB248426)

This data was developed using ab133251, the same antibody clone in a different buffer formulation.
Purified ab133251 at 1/20 dilution (0.6μg) immunoprecipitating c-Rel in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab133251 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab133251 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 78 kDa
Fresh lysate is required to avoid protein degradation.

All lanes:

Immunoprecipitation - Anti-c-Rel antibody [EPR2559(2)] (<a href='/en-us/products/primary-antibodies/c-rel-antibody-epr25592-ab133251'>ab133251</a>)

Predicted band size: 68 kDa

false

• WB

Lab

Western blot - Anti-c-Rel antibody [EPR2559(2)] - BSA and Azide free (AB248426)

This data was developed using ab133251, the same antibody clone in a different buffer formulation.

Western blot : Anti-c-Rel antibody [EPR2559(2)] ab133251 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 74 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in REL knockout HCT 116 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-c-Rel antibody [EPR2559(2)] (<a href='/en-us/products/primary-antibodies/c-rel-antibody-epr25592-ab133251'>ab133251</a>) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 at 20 µg

Lane 2:

Western blot - Human REL knockout HCT116 cell line (<a href='/en-us/products/cell-lines/human-rel-knockout-hct116-cell-line-ab286471'>ab286471</a>) at 20 µg

Lane 3:

HeLa at 20 µg

Lane 4:

PC-3 at 20 µg

Secondary

All lanes:

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

Predicted band size: 69 kDa

Observed band size: 74 kDa,37 kDa

false

• WB

Unknown

Western blot - Anti-c-Rel antibody [EPR2559(2)] - BSA and Azide free (AB248426)

This data was developed using ab133251, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-c-Rel antibody [EPR2559(2)] (<a href='/en-us/products/primary-antibodies/c-rel-antibody-epr25592-ab133251'>ab133251</a>) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 2:

HuT-78 (Human Sezary syndrome cutaneous T 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

Predicted band size: 68 kDa

false