Name: Anti-ZEB1 antibody [EPR17375]
Brand: RabMAb
SKU: ab203829
Rating: 3 (3 reviews)

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Western blot - 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

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

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: 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.
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.
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17375
Isotype

IgG
Research areas

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human ZEB1 protein (ab152728)

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
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	(2)	1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/250.

Notes
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.

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.
Target information above from: UniProt accession P37275 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 6935 Human
- Omim: 189909 Human
- SwissProt: P37275 Human
- Unigene: 124503 Human
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

Western blot - Anti-ZEB1 antibody [EPR17375] (ab203829)

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.
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.
Western blot - Anti-ZEB1 antibody [EPR17375] (ab203829)

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
Immunocytochemistry/ Immunofluorescence - Anti-ZEB1 antibody [EPR17375] (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.
Western blot - Anti-ZEB1 antibody [EPR17375] (ab203829)

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 (Intracellular) - Anti-ZEB1 antibody [EPR17375] (ab203829)

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.
Western blot - Anti-ZEB1 antibody [EPR17375] (ab203829)

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.
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.
Anti-ZEB1 antibody [EPR17375] (ab203829)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab203829? Please let us know so that we can cite the reference in this datasheet.

ab203829 has been referenced in 72 publications.

Li D et al. MicroRNA-375 represses tumor angiogenesis and reverses resistance to sorafenib in hepatocarcinoma. Cancer Gene Ther 28:126-140 (2021). PubMed: 32616906
Shen M et al. GLI3 Promotes Invasion and Predicts Poor Prognosis in Colorectal Cancer. Biomed Res Int 2021:8889986 (2021). PubMed: 33506047
Xiong Y et al. The VIM-AS1/miR-655/ZEB1 axis modulates bladder cancer cell metastasis by regulating epithelial-mesenchymal transition. Cancer Cell Int 21:233 (2021). PubMed: 33902589
Dong Y et al. miR-369 inhibits Liver Cancer progression by targeting ZEB1 pathway and predicts the prognosis of HCC patients. J Cancer 12:3067-3076 (2021). PubMed: 33854606
You JH et al. Mitochondrial pyruvate carrier 1 regulates ferroptosis in drug-tolerant persister head and neck cancer cells via epithelial-mesenchymal transition. Cancer Lett 507:40-54 (2021). PubMed: 33741422

View all Publications for this product

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1-3 of 3 Abreviews or Q&A

Application

ChIP

Sample

Human Cell lysate - nuclear (Mesenchymal cells)

Negative control

Background regions with no known ATAC-seq peaks or histone modifications.

Specification

Mesenchymal cells

Detection step

Real-time PCR

Type

Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Formaldehyde

Positive control

Known ZEB1 binding regions based on publicly available CHIP-seq data using different antibodies

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jan 25 2023

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (chondrocyte)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Sodium Citrate

Permeabilization

Specification

chondrocyte

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Fixative

zinc buffered formalin

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 09 2021

Application

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)

Sample

Human Tissue sections (Lung)

Antigen retrieval step

Heat mediated - Buffer/Enzyme Used: Citrate

Permeabilization

Specification

Lung

Blocking step

Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Fixative

Formaldehyde

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Sep 11 2019

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Specificity

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (72)

Customer reviews and Q&As

ChIP abreview for Anti-ZEB1 antibody [EPR17375]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ZEB1 antibody [EPR17375]

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-ZEB1 antibody [EPR17375]