Anti-Nrf2 antibody

• WB

Supplier Data

Western blot - Anti-Nrf2 antibody (AB137550)

Western blot : Anti-NFE2L2 antibody (ab137550) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab137550 was shown to bind specifically to NFE2L2. A band was observed at 85 kDa in wild-type A549 cell lysates with no signal observed at this size in NFE2L2 knockout cell line ab285359 (knockout cell lysate ab289682). To generate this image, wild-type and NFE2L2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % 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 IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Nrf2 antibody (ab137550) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

NFE2L2 knockout A549 cell lysate at 20 µg

Lane 2:

Western blot - Human NFE2L2 knockout A549 cell line[21] (<a href='/en-us/products/cell-lines/human-nfe2l2-knockout-a549-cell-line21-ab285359'>ab285359</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

Huh7 cell lysate at 20 µg

Predicted band size: 68 kDa

Observed band size: 85 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Nrf2 antibody (AB137550)

ab137550 at 5 μg immunoprecipitating Nrf2 in HepG2 whole cell extracts.

Lane 1 (-) : Ctrl IgG instead of ab137550 in HepG2 whole cell extracts.
Lane 2 (+) : ab137550 + HepG2 whole cell extracts.

For western blotting an anti-Rabbit IgG was used as a secondary reagent.

All lanes:

Immunoprecipitation - Anti-Nrf2 antibody (ab137550)

Predicted band size: 68 kDa

false

• ChIP

Supplier Data

ChIP - Anti-Nrf2 antibody (AB137550)

ChIP was performed with HepG2 chromatin extract and 5 μg of either normal rabbit IgG or Anti-Nrf2 antibody (ab137550). The precipitated DNA was detected by PCR with primer set targeting to GCLC gene locus.

• WB

Supplier Data

Western blot - Anti-Nrf2 antibody (AB137550)

7.5% gel.

Running condition : 80V, 15min; 140V, 40min.

Transfer condition : Semi-dry, 18 V, 60min (Nitrocellulose membrane)

Blocking condition : 5% non-fat milk in TBST, RT, 60min.

Primary antibody incubation : 4°C overnight.

Secondary antibody incubation : Room temperature for 1 hour.

Washing condition : 5 ml TBST, 4 x 5min.

ECL detection.

The observed MW (110kDa) is based on PMID : 22703241

All lanes:

Western blot - Anti-Nrf2 antibody (ab137550) at 1/1000 dilution

Lane 1:

Untreated (-) MDA-MB-231 nuclear extract at 30 µg

Lane 2:

MDA-MB-231 nuclear extract treated with 30 μM tBHQ for 4 hours (+) at 30 µg

Secondary

All lanes:

Rabbit IgG antibody (HRP) at 1/10000 dilution

Predicted band size: 68 kDa

Observed band size: 110 kDa

true

• WB

Supplier Data

Western blot - Anti-Nrf2 antibody (AB137550)

Samples were separated by 10% SDS-PAGE.

All lanes:

Western blot - Anti-Nrf2 antibody (ab137550) at 1/2000 dilution

Lane 1:

Untreated HepG2 whole cell extracts at 30 µg

Lane 2:

Treated HepG2 whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 69 kDa

false

• WB

Supplier Data

Western blot - Anti-Nrf2 antibody (AB137550)

5% SDS-PAGE

The observed MW (110kDa) is based on PMID : 22703241

All lanes:

Western blot - Anti-Nrf2 antibody (ab137550) at 1/1000 dilution

Lane 1:

Non-transfected (-) HEK-293T whole cell extracts at 30 µg

Lane 2:

NRF2-transfected (+, including 3xFlag-tag) HEK-293T whole cell extracts at 30 µg

Secondary

All lanes:

HRP-conjugated anti-rabbit IgG antibody

Predicted band size: 68 kDa

Observed band size: 110 kDa

false