Anti-SNAIL antibody [EPR21043] - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-SNAIL antibody [EPR21043] - BSA and Azide free (AB229701)

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 1/1000 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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216347).

All lanes:

Immunoprecipitation - Anti-SNAIL antibody [EPR21043] (<a href='/en-us/products/primary-antibodies/snail-antibody-epr21043-ab216347'>ab216347</a>)

Predicted band size: 29 kDa

Observed band size: 29 kDa

true

• WB

Lab

Western blot - Anti-SNAIL antibody [EPR21043] - BSA and Azide free (AB229701)

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.

All lanes:

Western blot - Anti-SNAIL antibody [EPR21043] (<a href='/en-us/products/primary-antibodies/snail-antibody-epr21043-ab216347'>ab216347</a>) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SNAI1 CRISPR-Cas9 edited HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human SNAI1 (SNAIL) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-snai1-snail-knockout-hela-cell-line-ab265963'>ab265963</a>)

Predicted band size: 29 kDa

Observed band size: 33 kDa

false

• WB

Supplier Data

Western blot - Anti-SNAIL antibody [EPR21043] - BSA and Azide free (AB229701)

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).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab216347).

All lanes:

Western blot - Anti-SNAIL antibody [EPR21043] - BSA and Azide free (ab229701) at 1/1000 dilution

Lane 1:

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

Lane 2:

HCT 116 (human colorectal carcinoma cell line) whole cell lysate at 20 µg

Lane 3:

MCF7 (human breast adenocarcinoma cell line) 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: 29 kDa

Observed band size: 29 kDa

true

Exposure time: 3min

• WB

Lab

Western blot - Anti-SNAIL antibody [EPR21043] - BSA and Azide free (AB229701)

This data was developed using the same antibody clone in a different buffer formulation (ab216347).

Western blot : Anti-SNAI1 antibody [EPR21043] (ab216347) staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab216347 was shown to bind specifically to SNAI1. A band was observed at 29 kDa in wild-type A549 cell lysates with no signal observed at this size in SNAI1 knockout cell line. To generate this image, wild-type and SNAI1 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-SNAIL antibody [EPR21043] (<a href='/en-us/products/primary-antibodies/snail-antibody-epr21043-ab216347'>ab216347</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

SNAI1 knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HeLa ab255929 cell lysate at 20 µg

Lane 4:

SNAI1 knockout HeLa <a href='/en-us/products/primary-antibodies/snail-antibody-epr21043-ab216347'>ab216347</a> cell lysate at 20 µg

Secondary

All lanes:

Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

Observed band size: 29 kDa

false