Recombinant Anti-SNAIL antibody [EPR21043] (ab216347)

False colour image of Western blot: Anti-SNAIL antibody [EPR21043] staining at 1/500 dilution, shown in green; loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) staining at 1/20000 dilution, shown in red. In Western blot, ab216347 was shown to bind specifically to SNAIL. A band was observed at 33 kDa in wild-type HeLa cell lysates with no signal observed at this size in SNAI1 CRISPR-Cas9 edited cell line ab265963 (CRISPR-Cas9 edited cell lysate ab257692). The band observed in the CRISPR-Cas9 edited lysate lane below 33 kDa is likely to represent a truncated form of SNAIL. 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 SNAI1 CRISPR-Cas9 edited HeLa 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

SNAIL was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab216347 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab216347 at 1/500 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab216347 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab216347 in HeLa whole cell lysate.

Exposure time: 10 seconds.

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

Blocking/Dilution buffer: 5% NFDM/TBST.

SNAIL expression is not detectable in MCF7 cells, which is consistent with what has been described in the literature (PMID: 10655587 and 22028892).

Blocking/Dilution buffer: 5% NFDM/TBST.