Anti-TCF12 antibody

• IP

Supplier Data

Immunoprecipitation - Anti-TCF12 antibody (AB245540)

TCF12 was immunoprecipitated from HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate (1 mg for IP, 20% of IP loaded) with ab245540 at 3 μg/mg lysate. Western blot was performed from the immunoprecipitate using ab245540 at 1 μg/ml.

Lane 1 : ab245540 IP in HeLa whole cell lysate.
Lane 2 : Control IgG IP in HeLa whole cell lysate.

Detection : Chemiluminescence with exposure time of 10 seconds.

All lanes:

Immunoprecipitation - Anti-TCF12 antibody (ab245540)

Predicted band size: 72 kDa

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• WB

Lab

Western blot - Anti-TCF12 antibody (AB245540)

Western blot : Anti-TCF12 antibody staining at 1/1000 dilution, shown in black; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab245540 was shown to bind specifically to TCF12. A band was observed at 85 kDa in wild-type Jurkat cell lysates with no signal observed at this size in TCF12 CRISPR-Cas9 edited cell line ab274931 (CRISPR-Cas9 edited cell lysate ab274989). The band observed in the CRISPR-Cas9 edited lysate lane below 85 kDa is likely to represent a truncated form of TCF12. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and TCF12 CRISPR-Cas9 edited Jurkat 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 before development with Optiblot (ECL reagent ab133456) and imaged with 20 minutes exposure time. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TCF12 antibody (ab245540) at 1/1000 dilution

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

TCF12 CRISPR-Cas9 edited Jurkat cell lysate at 20 µg

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Observed band size: 85 kDa

false

• WB

Lab

Western blot - Anti-TCF12 antibody (AB245540)

Western blot : Anti-TCF12 antibody ab245540 staining at 1/1000 dilution, shown in black; Mouse anti GAPDH (ab8245) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 80 kDa in Wild-type U-87 MG cell lysates with no signal observed at this size in TCF12 knockout U-87 MG 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 HRP (H+L) & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TCF12 antibody (ab245540) at 1/1000 dilution

Lane 1:

Wild-type U-87 MG at 20 µg

Lane 2:

Western blot - Human TCF12 knockout U-87 MG cell line (ab306824) at 20 µg

Lane 3:

HeLa at 20 µg

Secondary

Lanes 1 - 3:

Goat anti-Rabbit HRP (H+L) at 1/20000 dilution

Lanes 1 - 3:

Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 73 kDa

Observed band size: 80 kDa

true

Exposure time: 10s

• WB

Unknown

Western blot - Anti-TCF12 antibody (AB245540)

All lanes:

Western blot - Anti-TCF12 antibody (ab245540) at 0.1 µg/mL

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 50 µg

Lane 2:

HeLa whole cell lysate at 15 µg

Lane 3:

HeLa whole cell lysate at 5 µg

Predicted band size: 72 kDa

true

Exposure time: 3min