ZEB1 Antibody (639914)

ZEB1 was detected in immersion fixed MDA-MB-231 human breast cancer cell line using Mouse Anti-Human ZEB1 Monoclonal Antibody (Catalog # MAB6708) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red, upper panel; Catalog # NL007) and counterstained with DAPI (blue, lower panel). Specific staining was localized to nuclei and cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

MDA‑MB‑231 human breast cancer cell line was stained with Mouse Anti-Human ZEB1 Monoclonal Antibody (Catalog # MAB6708, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0101B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.

RAB25 is a ZEB1 target gene. (A) RAB25 expression positively correlated with E-cadherin but negatively with ZEB1 in a series of lung cancer cell lines. RNA expression was measured by quantitative real-time PCR in 22 NSCLC cell lines, two NHBE cultures, and two immortalized human airway primary cell lines (BAES2B and FC6625-2 3KT). Cells are ranked from left to right with increasing RAB25 mRNA level. Values are expressed as percent of the geometric mean between GAPDH and actin mRNA. The experiment was done twice with qRT-PCR in duplicate. (B) RAB25 mRNA level is decreased by ZEB1 overexpression in H358 FlipIn ZEB1 cells (left) and by TGF beta treatment in H358 EV control cells (right). Values are expressed as percent of GAPDH for three independent experiments with qRT-PCR in duplicate. Bars = SD. (C) Western blot: RAB25 protein level is decreased by ZEB1 overexpression during 1 to 5 days of DOX treatment in H358 FlipIn ZEB1cells. E-cadherin is decreased as well. Actin is the loading control. Protein molecular weights are indicated in kDa on the left. Image collected and cropped by CiteAb from the following publication (//pubmed.ncbi.nlm.nih.gov/24216980), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: Flow, CyTOF-ready, ICC/IF
• Clone: 639914
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

ZEB1 Antibody (639914) Summary

• Immunogen: E. coli-derived recombinant human ZEB1
  Glu430-Ser575
  Accession # P37275
• Specificity: Detects human ZEB1 in direct ELISAs.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: ZEB1
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Flow Cytometry 2.5 ug/10^6 cells
  • Immunocytochemistry 8-25 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for ZEB1 Antibody (639914)

• AREB6
• AREB6MGC133261
• BZP
• delta-crystallin enhancer binding factor 1
• DELTAEF1
• FECD6
• Negative regulator of IL2
• NIL-2-A zinc finger protein
• NIL2A
• NIL-2-A
• posterior polymorphous corneal dystrophy 3
• PPCD3
• TCF8
• TCF-8
• TCF8BZP
• transcription factor 8 (represses interleukin 2 expression)
• Transcription factor 8
• ZEB
• ZEB1
• ZFHEP
• ZFHX1A
• zinc finger E-box binding homeobox 1
• zinc finger E-box-binding homeobox 1
• zinc finger homeodomain enhancer-binding protein

Background

Zinc finger E-box-binding homeobox 1 (ZEB1; also transcription factor 8 (TCF-8)) is a 124 kDa member of the delta-EF1/ZFH-1 C2H2-type zinc finger family.  Human ZEB1 is 1124 amino acids (aa) in length. The protein contains seven C2H2-type zinc fingers and one homeobox DNA-binding domain. In addition, there are eight phosphoserines and one phosphothreonine. Residues 989-1124 make up a glutamine-rich area. Within aa 430-575, human ZEB1 shares 84% and 82% aa sequence identity with mouse and rat ZEB1, respectively. The protein is expressed in heart and skeletal muscle, and defects in ZEB1 are the cause of posterior polymorphous corneal dystrophy type 3, a rare disease involving metaplasia and overgrowth of the corneal endothelial cells.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for ZEB1 Antibody (MAB6708)(4)

Publications using MAB6708

• S Hasse, T Meder, E Freund, T von Woedtk, S Bekeschus Plasma Treatment Limits Human Melanoma Spheroid Growth and Metastasis Independent of the Ambient Gas Composition Cancers (Basel), 2020-09-09;12(9):. 2020-09-09 [PMID: 32917026] (Flow Cytometry, Human)
• P Lindner, S Paul, M Eckstein, C Hampel, JK Muenzner, K Erlenbach-, HP Ahmed, V Mahadevan, T Brabletz, A Hartmann, J Vera, R Schneider- EMT transcription factor ZEB1 alters the epigenetic landscape of colorectal cancer cells Cell Death Dis, 2020-02-24;11(2):147. 2020-02-24 [PMID: 32094334] (ICC, Human)
• Kinchen J, Chen HH, Parikh K et al. Structural Remodeling of the Human Colonic Mesenchyme in Inflammatory Bowel Disease. Cell 9/25/2018 [PMID: 30270042]
• Qinghua Ye, Lei Lei, Lingyun Shao, Jing Shi, Jun Jia, Xiaowen Tong MicroRNA-141 inhibits epithelial-mesenchymal transition, and ovarian cancer cell migration and invasion Molecular Medicine Reports 2017-09-13 [PMID: 28901523]

FAQs for ZEB1 Antibody (MAB6708). (Showing 1 - 1 of 1 FAQs).

1. In looking at the Zeb1 antibodies on your website, the molecular weights of Zeb1 detected by several antibodies are different. Why is this?
   • ZEB1 is a rather large protein and undergoes a lot of post translational modifications such as phosphorylation and glycosylation (please see: UniProt P37275). The reason band patterns are different in the images may be due to sample type and how highly modified the ZEB1 protein is that the antibody is detecting. The more post translational modifications the higher you will detect the protein.

Bioinformatics

• Gene Symbol: ZEB1
• Uniprot:
  • P37275()