Overview

  • Product name

    Anti-ZEB1 antibody [EPR17375]
    See all ZEB1 primary antibodies
  • Description

    Rabbit monoclonal [EPR17375] to ZEB1
  • Host species

    Rabbit
  • Specificity

    Expression levels of the target protein vary with sample type and some optimisation may be required.

  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human ZEB1 aa 1-150. The exact sequence is proprietary.
    Database link: P37275

  • Positive control

    • WB: HEK293, HAP1 and HeLa, MDA-MB231, SKOV3, MCF7, A549 cell lysates. Human Ovary lysates. IHC-P: Human cervix carcinoma and breast carcinoma tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab203829 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
WB 1/500. Detects a band of approximately 200 kDa (predicted molecular weight: 124 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.
Flow Cyt 1/50.

For unpurified use at 1/1000 dilution.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

Target

  • Function

    Inhibits interleukin-2 (IL-2) gene expression. May be responsible for transcriptional repression of the IL-2 gene. Enhances or represses the promoter activity of the ATP1A1 gene depending on the quantity of cDNA and on the cell type. Represses E-cadherin promoter and induces an epithelial-mesenchymal transition (EMT) by recruiting SMARCA4/BRG1. Represses BCL6 transcription in the presence of the corepressor CTBP1. Promotes tumorigenicity by repressing stemness-inhibiting microRNAs.
  • Tissue specificity

    Colocalizes with SMARCA4/BRG1 in E-cadherin-negative cells from established lines, and stroma of normal colon as well as in de-differentiated epithelial cells at the invasion front of colorectal carcinomas (at protein level). Expressed in heart and skeletal muscle, but not in liver, spleen, or pancreas.
  • Involvement in disease

    Defects in ZEB1 are the cause of posterior polymorphous corneal dystrophy type 3 (PPCD3) [MIM:609141]. PPCD is a rare disease involving metaplasia and overgrowth of corneal endothelial cells. In patients with PPCD, these cells manifest in an epithelial morphology and gene expression pattern, produce an aberrant basement membrane, and, sometimes, spread over the iris and nearby structures in a way that increases the risk for glaucoma.
    Defects in ZEB1 are the cause of corneal dystrophy Fuchs endothelial type 6 (FECD6) [MIM:613270]. It is an ocular disorder caused by loss of endothelium of the central cornea. It is characterized by focal wart-like guttata that arise from Descemet membrane and develop in the central cornea, epithelial blisters, reduced vision and pain. Descemet membrane is thickened by abnormal collagenous deposition.
  • Sequence similarities

    Belongs to the delta-EF1/ZFH-1 C2H2-type zinc-finger family.
    Contains 7 C2H2-type zinc fingers.
    Contains 1 homeobox DNA-binding domain.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • AREB 6 antibody
    • AREB6 antibody
    • BZP antibody
    • Delta crystallin enhancer binding factor 1 antibody
    • DELTA EF1 antibody
    • FECD6 antibody
    • MGC133261 antibody
    • Negative regulator of IL 2 antibody
    • Negative regulator of IL2 antibody
    • NIL 2 A antibody
    • NIL 2 A zinc finger protein antibody
    • NIL 2A antibody
    • NIL-2-A zinc finger protein antibody
    • NIL2A antibody
    • Posterior polymorphous corneal dystrophy 3 antibody
    • PPCD3 antibody
    • Represses interleukin 2 expression antibody
    • TCF 8 antibody
    • TCF-8 antibody
    • TCF8 antibody
    • Transcription factor 8 (represses interleukin 2 expression) antibody
    • Transcription factor 8 antibody
    • ZEB 1 antibody
    • ZEB antibody
    • ZEB1 antibody
    • ZEB1_HUMAN antibody
    • ZFHEP antibody
    • ZFHX 1A antibody
    • ZFHX1A antibody
    • Zinc finger E box binding homeobox 1 antibody
    • Zinc finger E-box-binding homeobox 1 antibody
    • Zinc finger homeodomain enhancer binding protein antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: ZEB1 knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)

    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. 

  • 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.

  • All lanes : Anti-ZEB1 antibody [EPR17375] (ab203829) at 1/500 dilution

    Lane 1 : MDA-MB231 (Human breast adenocarcinoma epithelial cell)whole cell lysates with 5% NFDM/TBST
    Lane 2 : SKOV3 (Human ovarian cancer epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 3 : MCF7 (Human breast adenocarcinoma epithelial cell)whole cell lysates with 5% NFDM/TBST
    Lane 4 : A549 (Human lung carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
    Lane 5 : Human Ovary lysates with 5% NFDM/TBST

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 124 kDa
    Observed band size: 200 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes
  • 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.

  • Anti-ZEB1 antibody [EPR17375] (ab203829) at 1/1000 dilution + HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 124 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?


    Exposure time: 15 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 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.

  • Anti-ZEB1 antibody [EPR17375] (ab203829) at 1/1000 dilution + HeLa (Human epithelial cell line from cervix adenocarcinoma) cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 124 kDa
    Observed band size: 200 kDa why is the actual band size different from the predicted?


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • 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.

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

References

This product has been referenced in:

  • Yang X & Meng T MicroRNA-431 affects trophoblast migration and invasion by targeting ZEB1 in preeclampsia. Gene 683:225-232 (2019). Read more (PubMed: 30315928) »
  • Jin H  et al. Long non-coding RNA MIAT competitively binds miR-150-5p to regulate ZEB1 expression in osteosarcoma. Oncol Lett 17:1229-1236 (2019). Read more (PubMed: 30655889) »
See all 22 Publications for this product

Customer reviews and Q&As

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
No
Specification
Lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Formaldehyde

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Verified customer

Submitted Sep 11 2019

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