Recombinant Anti-ZEB1 antibody [EPR17375] (ab203829)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR17375] to ZEB1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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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.
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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 (intra): HeLa cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR17375 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
1/50.
For unpurified use at 1/1000 dilution. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/500. Detects a band of approximately 200 kDa (predicted molecular weight: 124 kDa).
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IHC-P | (2) |
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ICC/IF |
1/250.
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Notes |
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Flow Cyt (Intra)
1/50. For unpurified use at 1/1000 dilution. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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. |
Target
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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
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Database links
- Entrez Gene: 6935 Human
- Omim: 189909 Human
- SwissProt: P37275 Human
- Unigene: 124503 Human
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Alternative names
- AREB 6 antibody
- AREB6 antibody
- BZP antibody
see all
Images
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZEB1 antibody [EPR17375] (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.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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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 Fluorr® 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.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ZEB1 antibody [EPR17375] (ab203829)
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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (87)
ab203829 has been referenced in 87 publications.
- Takamaru N et al. Association of PD-L1 and ZEB-1 expression patterns with clinicopathological characteristics and prognosis in oral squamous cell carcinoma. Oncol Lett 23:75 (2022). PubMed: 35111244
- Peng M et al. Programmed death-ligand 1 signaling and expression are reversible by lycopene via PI3K/AKT and Raf/MEK/ERK pathways in tongue squamous cell carcinoma. Genes Nutr 17:3 (2022). PubMed: 35164673
- Lan H et al. Pro-angiogenic role of ZEB1 in skin wound healing by upregulating VEGFA via microRNA-206 suppression. Exp Dermatol 31:1392-1401 (2022). PubMed: 35570385
- Wu J et al. NPTX1 inhibits pancreatic cancer cell proliferation and migration and enhances chemotherapy sensitivity by targeting RBM10. Oncol Lett 23:154 (2022). PubMed: 35836482
- Xie T et al. LHX2 facilitates the progression of nasopharyngeal carcinoma via activation of the FGF1/FGFR axis. Br J Cancer 127:1239-1253 (2022). PubMed: 35864158