重组Anti-SNAIL抗体[EPR21043]

• IP

Unknown

Immunoprecipitation - Anti-SNAIL antibody [EPR21043] (AB216347)

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.

All lanes:

Immunoprecipitation - Anti-SNAIL antibody [EPR21043] (ab216347)

Predicted band size: 29 kDa

Observed band size: 29 kDa

true

• WB

Lab

Western blot - 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.

All lanes:

Western blot - Anti-SNAIL antibody [EPR21043] (ab216347) 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='/products/cell-lines/human-snai1-snail-knockout-hela-cell-line-ab265963'>ab265963</a>)

Predicted band size: 29 kDa

Observed band size: 33 kDa

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• WB

Lab

Western blot - 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 knockout cell line ab265963 (knockout cell lysate ab257692). The band observed in the knockout 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 knockout 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] (ab216347) at 1/500 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

SNAI1 knockout HeLa cell lysate at 20 µg

Predicted band size: 29 kDa

Observed band size: 33 kDa

false

• WB

Lab

Western blot - Anti-SNAIL antibody [EPR21043] (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] (ab216347) 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 ab216347 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

• WB

Supplier Data

Western blot - Anti-SNAIL antibody [EPR21043] (AB216347)

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).

All lanes:

Western blot - Anti-SNAIL antibody [EPR21043] (ab216347) 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='/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

Supplier Data

Western blot - Anti-SNAIL antibody [EPR21043] (AB216347)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SNAIL antibody [EPR21043] (ab216347) at 1/10000 dilution

Lane 1:

His-tagged human SNAIL recombinant protein (aa1-264)

Lane 2:

His-tagged human Slug recombinant protein (aa21-268)

Lane 3:

His-tagged human Slug recombinant protein (aa1-110)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 29 kDa

Observed band size: 29 kDa

true

Exposure time: 5s

• WB

CiteAb

Western blot - Anti-SNAIL antibody [EPR21043] (AB216347)

SNAIL western blot using anti-SNAIL antibody [EPR21043] ab216347. Publication image and figure legend from Rudzińska, M., Parodi, A., et al., 2020, Cancers (Basel), PubMed 32455715.

ab216347 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab216347 please see the product overview.

Effects of the peptides on E-cadherin and SNAIL1 protein expression : (a,b) protein expression of E-cadherin and SNAIL1 in 769-P and (c,d) A498 cells after 24, 48, and 72 h of treatment with the peptides. Data are represented as mean ± SD of at least three replicates. Significance was calculated through one-way ANOVA, followed by Dunnet’s test.

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