ZEB1 - Antibodies - The Human Protein Atlas


	Antibody HPA027524	Antibody CAB058686	Antibody CAB079943

ANTIBODY INFORMATION
Provider	Atlas Antibodies Sigma-Aldrich	Atlas Antibodies	NCI-CPTAC
Product name	HPA027524	AMAb90510	CPTC-ZEB1-2
Host species	Rabbit	Mouse	Rabbit
Clonalityⁱ The antibodies are designated mAB for monoclonal and pAb for polyclonal.	pAb	mAb	mAb
Concentration	0.275 mg/ml	Not known	Not known
Purity	Affinity purified using the PrEST-antigen as affinity ligand	Protein A/G	Not known
Released in versionⁱ The release of the Human Protein Atlas in which the antibody was first published.	5.0	11.0	20.0
Referencesⁱ References to publications in which the antibody has been used.	49	3
Proper citation	Atlas Antibodies Cat#HPA027524, RRID:AB_1844977	Atlas Antibodies Cat#AMAb90510, RRID:AB_2665569	n/a
Validation summaryⁱ All assays through which the antibody has been validated. Assays&annotation provide a detailed description of the different assays. The pie-charts indicate degree of validation.	ICC IHC WB PA	N/A ICC IHC N/A WB N/A PA	ICC IHC WB N/A PA
IMMUNOCYTOCHEMISTRYⁱ Immunocytochemistry is used to validate the antibody staining and for assessing and validating the protein expression pattern in selected human cell lines.
Validationⁱ Results of validation by standard or enhanced validation. Standard validation is based on concordance with available experimental gene/protein characterization data in the UniProtKB/Swiss-Prot database. Standard validation results in scores Supported, Approved or Uncertain. Enhanced validation is performed using either siRNA knockdown, tagged GFP cell lines or independent antibodies. For the siRNA validation the decrease in antibody-based staining intensity upon target protein downregulation is evaluated. For the GFP validation the signal overlap between the antibody staining and the GFP-tagged protein is evaluated. For the independent antibodies validation the evaluation is based on comparison of the staining of two (or more) independent antibodies directed towards independent epitopes on the protein. For all cases except the siRNA validation, an image representative of the antibody staining pattern is shown. For the siRNA validation, a box plot of the results is shown.	Supportedⁱ Reliability scores for antibodies used in immunocytochemistry are set by comparing the staining pattern in cell lines with external experimental evidence for protein localization. The scores are termed Supported, Approved and Uncertain. The subcellular location is supported by literature. Immunofluorescent staining of human cell line U2OS shows localization to nucleoplasm.	N/A	Supportedⁱ Reliability scores for antibodies used in immunocytochemistry are set by comparing the staining pattern in cell lines with external experimental evidence for protein localization. The scores are termed Supported, Approved and Uncertain. The subcellular location is supported by literature. Immunofluorescent staining of human cell line U2OS shows localization to nucleoplasm and nucleoli.

Antibody dilution	Human assay: U-251MG fixed with PFA, dilution: 1:100 Human assay: U2OS fixed with PFA, dilution: 1:100		Human assay: A-549 fixed with PFA, dilution: 1:50 Human assay: Rh30 fixed with PFA, dilution: 1:50 Human assay: U2OS fixed with PFA, dilution: 1:50
IMMUNOHISTOCHEMISTRYⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain.
Validationⁱ Results of validation by standard or enhanced validation based on assessment of antibody performance in 44 normal tissues. Standard validation results in scores Supported, Approved or Uncertain. An image representative of the antibody staining pattern is shown. Enhanced validation results in the score Enhanced and includes two methods: Orthogonal validation and Independent antibody validation. For orthogonal validation, representative images of high and low expression are shown. For independent antibody validation, four images of each independent antibody are displayed.	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Endometrium 1 RNA expression: 37.5 nTPM LOW EXPRESSION Pancreas RNA expression: 3.7 nTPM	Enhanced - Orthogonal Antibody staining mainly consistent with RNA expression data across 44 tissues. HIGH EXPRESSION Endometrium 1 RNA expression: 37.5 nTPM LOW EXPRESSION Pancreas RNA expression: 3.7 nTPM	Supportedⁱ Immunohistochemistry is used for validating antibody reliability by assessing staining pattern in 44 normal tissues. Validation scores include Enhanced, Supported, Approved and Uncertain. Immunohistochemical staining of human smooth muscle shows strong nuclear positivity in smooth muscle cells. Smooth muscle

Retrievalⁱ Antigen retrieval is a method used to restore/retrieve the epitope (antibody bidning region) of the target protein, cross-linked, and thus masked, during tissue preserving fixative treatment of the tissues.	HIER pH6	HIER pH6	HIER pH6
Antibody dilution	1:600	1:1000	1:300
Literature conformityⁱ Conformance of the expression pattern with available gene/protein characterization data in scientific literature and data from bioinformatic predictions. UniProt is used as the main source of gene/protein characterization data and when relevant, available publications and other sources of information are researched in depth. Extensive or sufficient gene/protein data requires that there is evidence of existence on a protein level and that a substantial quantity of published experimental data is available from literature and public databases. Limited protein/gene characterization data does not require evidence of existence on a protein level and refers to genes for which only bioinformatic predictions and scarce published experimental data is available.	Consistent with extensive gene/protein characterization data.	Consistent with extensive gene/protein characterization data.	Partly consistent with extensive gene/protein characterization data.
RNA consistencyⁱ Consistency between immunohistochemistry data and consensus RNA levels is divided into five different categories: i) High consistency, ii) Medium consistency, iii) Low consistency, iv) Very low consistency, and v) Cannot be evaluated.	High consistency between antibody staining and RNA expression data.	High consistency between antibody staining and RNA expression data.	Low consistency between antibody staining and RNA expression data.
WESTERN BLOTⁱ A Western blot analysis is performed on a panel of human tissues and cell lines to evaluate antibody specificity. For antibodies with unreliable result a revalidation using an over-expression lysate is performed.
Validationⁱ Western Blot is used for quality control of the polyclonal antibodies generated in the project. After purification, the antibodies are used to detect bands in a setup of lysate and different tissues. The result is then scored Enhanced, Supported, Approved, or Uncertain. Enhanced validation includes five different methods: Genetic validation, Recombinant expression validation, Independent antibody validation, Orthogonal validation and Capture MS validation.	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. 230 130 95 72 56 36 28 17 11	Uncertainⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Weak band of predicted size but with additional bands of higher intensity also present. Analysis performed using a standard panel of samples.	Supportedⁱ The staining of an antibody is evaluated by Western Blot through analysis of samples from different cell lysates. A supportive score is given if band(s) of predicted size in kDa (+/-20%) is detected. Band of predicted size in kDa (+/-20%) with additional bands present. Analysis performed using a standard panel of samples. 250 130 100 70 55 35 25 15 10

Antibody dilution	1:250	1:500	1:500
PROTEIN ARRAY
Validationⁱ A protein array containing 384 different antigens including the antibody target is used to analyse antibody specificity. Depending on the array interaction profile the antibody is scored as Supported, Approved, or Uncertain.	Supported Pass with single peak corresponding to interaction only with its own antigen. Antibody specificity analysis with protein arrays. Predicted and matching interactions are shown in green.	N/A	N/A

Antibody dilution	1:3000
RELEVANT PUBLICATIONS
	Overexpression of ZEB1 associated with metastasis and invasion in patients with gastric carcinoma Jia B et al Mol Cell Biochem 2012;366(1-2):223-9	Grainyhead-like 2 (GRHL2) distribution reveals novel pathophysiological differences between human idiopathic pulmonary fibrosis and mouse models of pulmonary fibrosis Varma S et al Am J Physiol Lung Cell Mol Physiol 2014;306(5):L405-19 Application: ICC-IF
	Quantitative analysis of microRNAs in tissue microarrays by in situ hybridization Hanna JA et al Biotechniques 2012;52(4):235-45	Next-generation sequencing reveals novel differentially regulated mRNAs, lncRNAs, miRNAs, sdRNAs and a piRNA in pancreatic cancer Müller S et al Mol Cancer 2015;14:94 Application: IHC
	Poor-prognosis colon cancer is defined by a molecularly distinct subtype and develops from serrated precursor lesions De Sousa E Melo F et al Nat Med 2013;19(5):614-8	Serglycin as a potential biomarker for glioma: association of serglycin expression, extent of mast cell recruitment and glioblastoma progression Roy A et al Oncotarget 2017;8(15):24815-24827 Application: IHC
	Zeb1 regulates E-cadherin and Epcam (epithelial cell adhesion molecule) expression to control cell behavior in early zebrafish development Vannier C et al J Biol Chem 2013;288(26):18643-59
	The microRNA-200 family targets multiple non-small cell lung cancer prognostic markers in H1299 cells and BEAS-2B cells Pacurari M et al Int J Oncol 2013;43(2):548-60
	Epithelial to mesenchymal transition in murine tracheal allotransplantation: an immunohistochemical observation Konoeda C et al Transplant Proc 2013;45(5):1797-801
	Dual roles of the transcription factor grainyhead-like 2 (GRHL2) in breast cancer Werner S et al J Biol Chem 2013;288(32):22993-3008 Application: ChIP, WB
	Direct RNA sequencing mediated identification of mRNA localized in protrusions of human MDA-MB-231 metastatic breast cancer cells Jakobsen KR et al J Mol Signal 2013;8(1):9
	Activation of miR200 by c-Myb depends on ZEB1 expression and miR200 promoter methylation Pieraccioli M et al Cell Cycle 2013;12(14):2309-20 Application: WB
	Transcription factors OVOL1 and OVOL2 induce the mesenchymal to epithelial transition in human cancer Roca H et al PLoS One 2013;8(10):e76773
	Epithelial-mesenchymal transition induces endoplasmic-reticulum-stress response in human colorectal tumor cells Zeindl-Eberhart E et al PLoS One 2014;9(1):e87386
	CD95 and CD95L promote and protect cancer stem cells Ceppi P et al Nat Commun 2014;5:5238 Application: ICC-IF, WB
	The clinical role of epithelial-mesenchymal transition and stem cell markers in advanced-stage ovarian serous carcinoma effusions Davidson B et al Hum Pathol 2015;46(1):1-8 Application: IHC
	ZEB1-associated drug resistance in cancer cells is reversed by the class I HDAC inhibitor mocetinostat Meidhof S et al EMBO Mol Med 2015;7(6):831-47 Application: ICC-IF, IHC, WB
	The EMT-activator ZEB1 induces bone metastasis associated genes including BMP-inhibitors Mock K et al Oncotarget 2015;6(16):14399-412 Application: WB
	EMT-associated factors promote invasive properties of uveal melanoma cells Asnaghi L et al Mol Vis 2015;21:919-29 Application: WB
	The ZEB1/miR-200c feedback loop regulates invasion via actin interacting proteins MYLK and TKS5 Sundararajan V et al Oncotarget 2015;6(29):27083-96 Application: WB
	Low levels of Caspase-3 predict favourable response to 5FU-based chemotherapy in advanced colorectal cancer: Caspase-3 inhibition as a therapeutic approach Flanagan L et al Cell Death Dis 2016;7:e2087 Application: IHC
	Expression of zinc finger E-box-binding homeobox factor 1 in epithelial ovarian cancer: A clinicopathological analysis of 238 patients Li X et al Mol Clin Oncol 2016;4(1):18-22 Application: IHC
	ZEB1 turns into a transcriptional activator by interacting with YAP1 in aggressive cancer types Lehmann W et al Nat Commun 2016;7:10498 Application: IHC, WB
	Zeb1 controls neuron differentiation and germinal zone exit by a mesenchymal-epithelial-like transition Singh S et al Elife 2016;5: Application: IP
	TGFβ signaling directs serrated adenomas to the mesenchymal colorectal cancer subtype Fessler E et al EMBO Mol Med 2016;8(7):745-60 Application: IHC
	Radiation driven epithelial-mesenchymal transition is mediated by Notch signaling in breast cancer Kim RK et al Oncotarget 2016;7(33):53430-53442 Application: ICC-IF
	Loss of ZFP36 expression in colorectal cancer correlates to wnt/ ß-catenin activity and enhances epithelial-to-mesenchymal transition through upregulation of ZEB1, SOX9 and MACC1 Montorsi L et al Oncotarget 2016;7(37):59144-59157 Application: WB
	Reciprocal regulation of the cholinic phenotype and epithelial-mesenchymal transition in glioblastoma cells Koch K et al Oncotarget 2016;7(45):73414-73431 Application: WB
	Resetting cancer stem cell regulatory nodes upon MYC inhibition Galardi S et al EMBO Rep 2016;17(12):1872-1889 Application: WB
	A novel ZEB1/HAS2 positive feedback loop promotes EMT in breast cancer Preca BT et al Oncotarget 2017;8(7):11530-11543 Application: IHC, WB
	Succession of transiently active tumor-initiating cell clones in human pancreatic cancer xenografts Ball CR et al EMBO Mol Med 2017;9(7):918-932 Application: ICC-IF
	Loss of DIP2C in RKO cells stimulates changes in DNA methylation and epithelial-mesenchymal transition Larsson C et al BMC Cancer 2017;17(1):487 Application: WB
	Cellular heterogeneity contributes to subtype-specific expression of ZEB1 in human glioblastoma Euskirchen P et al PLoS One 2017;12(9):e0185376 Application: ICC-IF, IHC
	Phytosphingosine exhibits an anti-epithelial-mesenchymal transition function by the inhibition of EGFR signaling in human breast cancer cells Kang HM et al Oncotarget 2017;8(44):77794-77808 Application: ICC-IF
	ADHFE1 is a breast cancer oncogene and induces metabolic reprogramming Mishra P et al J Clin Invest 2018;128(1):323-340 Application: WB
	Distinguishing mechanisms underlying EMT tristability Jia D et al Cancer Converg 2017;1(1):2 Application: ICC-IF
	Caspase-independent cell death does not elicit a proliferative response in melanoma cancer cells Roumane A et al BMC Cell Biol 2018;19(1):11 Application: WB
	PAR1 signaling on tumor cells limits tumor growth by maintaining a mesenchymal phenotype in pancreatic cancer Tekin C et al Oncotarget 2018;9(62):32010-32023 Application: ICC-IF
	Clinical Impact of Epithelial-to-Mesenchymal Transition Regulating MicroRNAs in Pancreatic Ductal Adenocarcinoma Dhayat SA et al Cancers (Basel) 2018;10(9): Application: IHC
	EMT induced by loss of LKB1 promotes migration and invasion of liver cancer cells through ZEB1-induced YAP signaling Qiu B et al Oncol Lett 2018;16(5):6465-6471 Application: WB
	Tumor-associated macrophages (TAMs) depend on ZEB1 for their cancer-promoting roles Cortés M et al EMBO J 2017;36(22):3336-3355 Application: IHC
	Regulation of muscle atrophy-related genes by the opposing transcriptional activities of ZEB1/CtBP and FOXO3 Ninfali C et al Nucleic Acids Res 2018;46(20):10697-10708 Application: WB
	The Tumor Suppressor SASH1 Interacts With the Signal Adaptor CRKL to Inhibit Epithelial-Mesenchymal Transition and Metastasis in Colorectal Cancer Franke FC et al Cell Mol Gastroenterol Hepatol 2019;7(1):33-53 Application: WB
	Carcinosarcoma of the esophagus: A report of 6 cases associated with zinc finger E-box-binding homeobox 1 expression Harada H et al Oncol Lett 2019;17(1):578-586 Application: IHC
	Early-stage serrated adenocarcinomas are divided into several molecularly distinct subtypes Hirano D et al PLoS One 2019;14(2):e0211477 Application: IHC
	ZEB1 protects skeletal muscle from damage and is required for its regeneration Siles L et al Nat Commun 2019;10(1):1364 Application: ICC-IF, WB
	Interconnected feedback loops among ESRP1, HAS2, and CD44 regulate epithelial-mesenchymal plasticity in cancer Jolly MK et al APL Bioeng 2018;2(3):031908 Application: WB
	ACVR1C/SMAD2 signaling promotes invasion and growth in retinoblastoma Asnaghi L et al Oncogene 2019;38(12):2056-2075 Application: WB
	Downregulation of Nodal inhibits metastatic progression in retinoblastoma Asnaghi L et al Acta Neuropathol Commun 2019;7(1):137 Application: WB
	Extracellular vesicle cross-talk between pulmonary artery smooth muscle cells and endothelium during excessive TGF-β signalling: implications for PAH vascular remodelling de la Cuesta F et al Cell Commun Signal 2019;17(1):143 Application: ICC-IF
	Hypoxia-induced HIF-1α and ZEB1 are critical for the malignant transformation of ameloblastoma via TGF-β-dependent EMT Yoshimoto S et al Cancer Med 2019;8(18):7822-7832 Application: ICC-IF, IHC, WB
	Cancer-associated stroma significantly contributes to the mesenchymal subtype signature of serous ovarian cancer Zhang Q et al Gynecol Oncol 2019;152(2):368-374 Application: IHC
ANTIGEN INFORMATION
Antigen	Recombinant protein fragment	Recombinant protein	Recombinant protein
Length (aa)	130
Antigen sequence	`EAEKPESSVSSATGDGNLSPSQPPLKNLLSLLKAYYALNAQPSAEELSKI ADSVNLPLDVVKKWFEKMQAGQISVQSSEPSSPEPGKVNIPAKNNDQPQS ANANEPQDSTVNLQSPLKMTNSPVLPVGST`
Matching transcripts	ZEB1-201 - ENSP00000319248 [100%] ZEB1-202 - ENSP00000415961 [100%] ZEB1-204 - ENSP00000391612 [100%] ZEB1-207 - ENSP00000444891 [100%] ZEB1-217 - ENSP00000452787 [100%]
Matching mouse transcripts	ENSMUSP00000025081 [74%] ENSMUSP00000028229 [28%] ENSMUSP00000158253 [28%]
ANTIGEN VIEWⁱ The Structure section provides in-house generated structures, predicted using the Alphafold source code, for the majority of the proteins and their related isoforms. Displaying protein features on the AlphaFold structures Individual splice variants can be selected in the top part of the Protein Browser (see below) and different transcript-related features such as transmembrane regions, InterPro domains and antigen sequences for antibodies can be displayed in the structure by clicking on the respective features in the Protein Browser. Clinical and population-based amino acid variants based on data from the Ensembl variation database and AlphaMissense (AM) predictions can be highlighted using the sliders to the right of the structure. These can also be used to colour the entire structure by residue index or make the structure autorotate.The structures are displayed using the NGL Viewer and can also be zoomed-in and rotated manually. The Protein Browser The ProteinBrowser displays the antigen location on the target protein(s) and the features of the target protein. Transcript names and schematic transcript structures including exons, introns and UTRs for the different isoforms are shown on top, and can be used to switch between the structures for the different splice variants. At the top of the view, the position of the antigen (identified by the corresponding HPA identifier) is shown as a green bar. A yellow triangle on the bar indicates a <100% sequence identity to the protein target. Below the antigens, the maximum percent sequence identity of the protein to all other proteins from other human genes is displayed, using a sliding window of 10 aa residues (HsID 10) or 50 aa residues (HsID 50). The region with the lowest possible identity is always selected for antigen design, with a maximum identity of 60% allowed for designing a single-target antigen (read more). The curve in blue displays the predicted antigenicity i.e. the tendency for different regions of the protein to generate an immune response, with peak regions being predicted to be more antigenic.The curve shows average values based on a sliding window approach using an in-house propensity scale. (read more). Signal peptides (turquoise) and membrane regions (orange) based on predictions using the majority decision methods MDM and MDSEC are also displayed. Low complexity regions are shown in yellow and InterPro regions in green. Common (purple) and unique (grey) regions between different splice variants of the gene are also displayed (read more), and at the bottom of the protein view is the protein scale.

ZEB1-201 ZEB1-202 ZEB1-204 ZEB1-207 ZEB1-217

Displaying protein features on the AlphaFold structures

The Protein Browser