Anti-ZEB1 antibody [EPR17375] - BSA and Azide free

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (AB228986)

Intracellular flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ZEB1 with purified ab203829 at 1/50 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203829).

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (AB228986)

Intracellular Flow Cytometry analysis of HeLa cells labelling ZEB1 with ab203829 at 1/1000 (red). Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. An Alexa Fluor^� 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203829).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (AB228986)

This IHC data was generated using the same anti-ZEB1 antibody clone, EPR17375, in a different buffer formulation (cat# ab203829).

Immunohistochemical analysis of paraffin-embedded Human cervix carcinoma tissue labeling ZEB1 with ab203829 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human cervix carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (AB228986)

This ICC data was generated using the same anti-ZEB1 antibody clone, EPR17375, in a different buffer formulation (cat# ab203829).

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling ZEB1 with ab203829 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows :
-ve control 1 : ab203829 at 1/250 dilution followed by ab150120 (Alexa Fluor^® 594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor^® 488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (AB228986)

Immunohistochemical analysis of paraffin-embedded Human breast carcinoma tissue labeling ZEB1 with ab203829 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on Human breast carcinoma tissue is observed. Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203829).

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Lab

Western blot - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (AB228986)

**Lanes 1 - 3 : ** Merged signal (red and green). Green - ab203829 observed at 200 kDa. Red - loading control, ab9484, observed at 37 kDa. ab203829 was shown to specifically react with ZEB1 in wild-type HAP1 cells as signal was lost in ZEB1 knockout cells. Wild-type and ZEB1 knockout samples were subjected to SDS-PAGE. ab203829 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging. This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab203829).

Lane 1:

Western blot - Anti-ZEB1 antibody [EPR17375] (<a href='/en-us/products/primary-antibodies/zeb1-antibody-epr17375-ab203829'>ab203829</a>)

Lanes 2 - 3:

Western blot - Anti-ZEB1 antibody [EPR17375] - BSA and Azide free (ab228986) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

ZEB1 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 124 kDa

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