Anti-TCF12 antibody [7B3]

• WB

Lab

Western blot - Anti-TCF12 antibody [7B3] (AB243149)

Western blot : Anti-TCF12 antibody [7B3] staining at 0.394 ug/ml, shown in black; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab243149 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-Rat (H+L) and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-TCF12 antibody [7B3] (ab243149) at 0.394 µg/mL

Lane 1:

Wild-type Jurkat cell lysate at 20 µg

Lane 2:

TCF12 CRISPR-Cas9 edited Jurkat cell lysate at 20 µg

Lane 2:

Western blot - Human TCF12 knockout Jurkat cell line (<a href='/en-us/products/cell-lines/human-tcf12-knockout-jurkat-cell-line-ab274931'>ab274931</a>)

Lane 2:

Western blot - Human TCF12 knockout Jurkat cell lysate (ab274989)

Lane 3:

HeLa cell lysate at 20 µg

Lane 4:

MOLT-4 cell lysate at 20 µg

Predicted band size: 72 kDa

Observed band size: 85 kDa

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• WB

Supplier Data

Western blot - Anti-TCF12 antibody [7B3] (AB243149)

The expression molecular weight observed is consistent with what has been described in the literature (PMID : 16785505).

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

All lanes:

Western blot - Anti-TCF12 antibody [7B3] (ab243149) at 0.394 µg/mL

Lane 1:

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

Lane 2:

SW620 (human colorectal adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

Raji (human Burkitt’s lymphoma B lymphocyte), whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte), whole cell lysate at 20 µg

Lane 5:

WEHI-231 (mouse B cell lymphoma B lymphocyte), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rat IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rat-igg-h-l-hrp-ab205720'>ab205720</a>) at 1/5000 dilution

Predicted band size: 72 kDa

Observed band size: 92 kDa

false

• WB

Lab

Western blot - Anti-TCF12 antibody [7B3] (AB243149)

Western blot : Rat monoclonal to TCF12 [7B3] ab243149 staining at 0.394 µg/mL, shown in green; Rabbit anti GAPDH (ab181602) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 80 kDa in Wild-type A549 cell lysates with no signal observed at this size in TCF12 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 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-Rat 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-TCF12 antibody [7B3] (ab243149) at 0.394 µg/mL

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

TCF12 knockout A549 at 20 µg

Secondary

Lanes 1 - 2:

Goat anti-Rat 800CW at 1/20000 dilution

Lanes 1 - 2:

Goat anti-Rabbit 680RD at 1/20000 dilution

Predicted band size: 80 kDa

Observed band size: 80 kDa,37 kDa

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