ZEB1 Antibody

Immunohistochemistry: ZEB1 Antibody [NBP1-05987] - Immunohistochemical staining results of EMT regulators in HG-PanIN: SNAIL, SLUG, TWIST1, and ZEB1. We categorized and defined three grades: grade 0, <10% positive staining; grade 1, 10%-50% positive; grade 2, 50%< positive Image collected and cropped by CiteAb from the following publication (//onlinelibrary.wiley.com/doi/abs/10.1002/cam4.2016) licensed under a CC-BY licence.

Western Blot: ZEB1 Antibody [NBP1-05987] - Samples: Whole cell lysate (15 ug) from HeLa, HEK293T, Jurkat, mouse TCMK-1, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-ZEB1 antibody (NBP1-05987) used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.

Immunohistochemistry-Paraffin: ZEB1 Antibody [NBP1-05987] - Sample: FFPE section of human breast carcinoma. Antibody: Affinity purified rabbit anti-ZEB1 (NBP1-05987) used at a dilution of 1:1000 (0.2 ug/ml). Detection: DAB.

Western Blot: ZEB1 Antibody [NBP1-05987] - Analysis of ZEB1 in transfected and native mouse cell lysate. Image courtesy of anonymous customer review.

Immunohistochemistry-Paraffin: ZEB1 Antibody [NBP1-05987] - Analysis of ZEB1 in human liver and spleen tissues. Image courtesy of anonymous customer review.

Immunohistochemistry-Paraffin: ZEB1 Antibody [NBP1-05987] - Sample: FFPE section of human breast carcinoma. Antibody: Affinity purified rabbit anti-ZEB1 (NBP1-05987) used at a dilution of 1:1000 (0.2 ug/ml). Detection: DAB.

Immunoprecipitation: ZEB1 Antibody [NBP1-05987] - Detection of human ZEB1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-ZEB1 antibody NBP1-05987 used for IP at 3 ug per reaction. ZEB1 was also immunoprecipitated by another rabbit anti-ZEB1 antibody. For blotting immunoprecipitated ZEB1, NBP1-05987 was used at 0.4 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, GS, ICC/IF, IHC, IHC-P, IP, MiAr, ChIP, KD, Single-Cell Western
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Concentration: 0.2 mg/ml

ZEB1 Antibody Summary

• Immunogen: The immunogen recognized by this antibody maps to a region between residue 1074 and 1124 of human zinc finger E-box binding homeobox 1 using the numbering given in entry NP_110378.3
• Marker: Mesenchymal Cells Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: ZEB1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Western Blot 1:2000-1:10000
  • Simple Western 1:50
  • Gel Super Shift Assays 1:1-1:100
  • Immunocytochemistry/Immunofluorescence 1:50-1:500
  • Immunohistochemistry 1:200-1:1000
  • Immunohistochemistry-Paraffin 1:200-1:1000
  • Immunoprecipitation 2-5 ug/mg lysate
  • Microarray
  • Chromatin Immunoprecipitation (ChIP)
  • Knockdown Validated
  • Single Cell Western 100 ug/ml
• Application Notes: Customers have also reported success in IHC on paraffin tissues, following antigen retrieval with citrate buffer. Gel Super Shift Assays (PMID: 21771782). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue. ICC/IF reactivity reported in scientific literature (PMID: 24334458). Use in microarray reported in scientific literature (PMID: 28955722).Use in chromatin immunoprecipitation reported in scientific literature (PMID: 29744893). ZEB1antibody validated for IHC-P, WB, IHC from verified customer reviews. Knockdown validation (PMID: 31776338).
• Control:

  ZEB1 Overexpression Lysate

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 28783105).

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: TBS and 0.1% BSA
• Preservative: 0.09% Sodium Azide
• Concentration: 0.2 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for ZEB1 Antibody

• 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

ZEB1 encodes a zinc finger transcription factor that represses T-lymphocyte-specific IL2 gene (MIM 147680) expression by binding to a negative regulatory domain 100 nucleotides 5-prime of the IL2 transcription start site (Williams et al., 1991 [PubMed 1840704]).[supplied by OMIM]

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 (NBP1-05987)(62)

Publications using NBP1-05987

• Wilkinson ML, Abramova E, Guo C et al. Fatty acid nitroalkenes inhibit the inflammatory response to bleomycin-mediated lung injury Toxicology and applied pharmacology Sep 12 2020 [PMID: 32931793] (SW, Mouse)
• Sato K, Hikita H, Myojin Y et al. Hyperglycemia enhances pancreatic cancer progression accompanied by elevations in phosphorylated STAT3 and MYC levels PLoS ONE Jul 1 2020 [PMID: 32609742] (WB, Mouse, Human)
• Huang C, Yoon C, Zhou XH et al. ERK1/2-Nanog signaling pathway enhances CD44(+) cancer stem-like cell phenotypes and epithelial-to-mesenchymal transition in head and neck squamous cell carcinomas Cell Death Dis Apr 23 2020 [PMID: 32327629] (WB, Human)
• Horn La, Riskin J, Hempel Ha et Al. Simultaneous inhibition of CXCR1/2, TGF-beta, and PD-L1 remodels the tumor and its microenvironment to drive antitumor immunity J Immunother Cancer Mar 1 2020 [PMID: 32188703] (IF, Human, Mouse)
• Ran J, Yang Hh, Huang Hp et Al. ZEB1 modulates endometrial receptivity through epithelial-mesenchymal transition in endometrial epithelial cells in vitro Biochem. Biophys. Res. Commun. Mar 2 2020 [PMID: 32139122] (WB, Human)
• Murata M, Ito T, Tanaka Y et al. OVOL2-Mediated ZEB1 Downregulation May Prevent Promotion of Actinic Keratosis to Cutaneous Squamous Cell Carcinoma J Clin Med Feb 25 2020 [PMID: 32106476] (WB, Human)
• High expression of ZEB1 in endometriosis and its role in 17b-estradiol-induced epithelial-mesenchymal transition. Wu R, Chen Z, Zhou W et al. Int J Clin Exp Pathol [PMID: 31949550] (IHC, Human)
• Miro C, Di Cicco E, Ambrosio R et al. Thyroid hormone induces progression and invasiveness of squamous cell carcinomas by promoting a ZEB-1/E-cadherin switch Nat Commun Nov 27 2019 [PMID: 31776338] (IHC, KD, Human, Mouse)
• Osada A H, Endo K et al. Addiction of mesenchymal phenotypes on the FGF/FGFR axis in oral squamous cell carcinoma cells. PLoS One May 11 2019 [PMID: 31682640] (WB, Human)
• Viswanathan K, Borczuk A The Diagnostic Utility of Zinc E-box 1 (ZEB1) Transcription Factor for Identification of Pulmonary Sarcomatoid Carcinoma in Cytologic and Surgical Specimens Journal of the American Society of Cytopathology Sep 1 2019 [PMID: 31677963] (IHC-P, Human)
• Hah YS, Cho HY, Jo SY et al. Nicotinamide N methyltransferase induces the proliferation and invasion of squamous cell carcinoma cells Oncol. Rep. Sep 16 2019 [PMID: 31545452] (WB, Human)
• Yoon C, Till J, Cho SJ, et al. KRAS activation in gastric adenocarcinoma stimulates epithelial-to-mesenchymal transition to cancer stem-like cells and promotes metastasis Mol. Cancer Res. 2019 Jun 19 [PMID: 31217166] (WB, Human)
• Sippel TR, Johnson AM, Li HY et al. Activation of PPAR gamma in Myeloid Cells Promotes Progression of Epithelial Lung Tumors Through TGF-beta 1 Mol. Cancer Res. May 14 2019 [PMID: 31088909] (WB, Mouse)
• Ordonez, LD;Hay, T;McEwen, R;Polanska, UM;Hughes, A;Delpuech, O;Cadogan, E;Powell, S;Dry, J;Tornillo, G;Silcock, L;Leo, E;O Connor, MJ;Clarke, AR;Smalley, MJ; Rapid activation of epithelial-mesenchymal transition drives PARP inhibitor resistance in Brca2-mutant mammary tumours Oncotarget Apr 16 2019 [PMID: 31080552] (WB, Mouse)
• Rios AC, Capaldo BD, Vaillant F et al. Intraclonal Plasticity in Mammary Tumors Revealed through Large-Scale Single-Cell Resolution 3D Imaging Cancer Cell Mar 14 2019 [PMID: 30930118] (ICC/IF, Mouse)
• Katsuno Y, Meyer DS, Zhang Z et al. Chronic TGF-b exposure drives stabilized EMT, tumor stemness, and cancer drug resistance with vulnerability to bitopic mTOR inhibition Sci Signal Feb 26 2019 [PMID: 30808819] (WB, Human)
• Fujiwara S, Saiki Y, Ishizawa K et al. Expression of SNAIL in accompanying PanIN is a key prognostic indicator in pancreatic ductal adenocarcinomas Cancer Med Feb 21 2019 [PMID: 30791220] (IHC-P, Human)
• Jia D, Augert A, Kim DW et al. Crebbp loss drives small cell lung cancer and increases sensitivity to HDAC inhibition. Cancer Discov Sep 4 2018 [PMID: 30181244] (IHC-P, Mouse)
• Knaack H, Lenk L, Philipp LM et al. Liver metastasis of pancreatic cancer: the hepatic microenvironment impacts differentiation and self-renewal capacity of pancreatic ductal epithelial cells Oncotarget Aug 3 2018 [PMID: 30167093] (WB, Mouse)
• Chen Y, LeBleu VS, Carstens JL et al. Dual reporter genetic mouse models of pancreatic cancer identify an epithelial-to-mesenchymal transition-independent metastasis program EMBO Mol Med Aug 17 2018 [PMID: 30120146] (ICC/IF, IHC-P, Mouse)
• Weng CH, Chen LY, Lin YC et al. Epithelial-mesenchymal transition (EMT) beyond EGFR mutations per se is a common mechanism for acquired resistance to EGFR TKI Oncogene Aug 15 2018 [PMID: 30111817] (WB, Human)
• Cho SJ, Yoon C, Lee JH et al. KMT2C mutations in diffuse type gastric adenocarcinoma promote epithelial-to-mesenchymal transition Clin. Cancer Res. Aug 14 2018 [PMID: 30108106] (WB, Human)
• Suzuki K, Kawataki T, Endo K et al. Expression of ZEBs in gliomas is associated with invasive properties and histopathological grade Oncol Lett. [PMID: 30008863] (IHC, Human)
• Chang KK, Yoon C, Yi BC et al. Platelet-derived growth factor receptor-alpha and-b promote cancer stem cell phenotypes in sarcomas Oncogenesis Jun 19 2018 [PMID: 29915281] (WB, Human)
• Maturi V, Enroth S, Heldin CH, Moustakas A. Genome-wide binding of transcription factor ZEB1 in triple-negative breast cancer cells. J. Cell. Physiol. Oct 1 2018 [PMID: 29744893] (ChIP, Human)
• Maturi V, Moren A, Enroth S et al. Genomewide binding of transcription factor Snail1 in triple-negative breast cancer cells. Mol Oncol Jun 1 2018 [PMID: 29729076] (WB, Human)
• Acharya KD, Nettles SA, Sellers KJ et al. The Progestin Receptor Interactome in the Female Mouse Hypothalamus: Interactions with Synaptic Proteins Are Isoform Specific and Ligand Dependent eNeuro 2017 Sep 28 [PMID: 28955722] (MiAr, Mouse)
• Lutz G, Jurak I, Kim ET et al. Viral Ubiquitin Ligase Stimulates Selective Host MicroRNA Expression by Targeting ZEB Transcriptional Repressors Viruses 2017 Aug 07 [PMID: 28783105] (WB, Rat)
• Liang X, Yuan X, Yu J et al. Histone Chaperone ASF1A Predicts Poor Outcomes for Patients With Gastrointestinal Cancer and Drives Cancer Progression by Stimulating Transcription of beta-Catenin Target Genes EBioMedicine. 2017 Jun 07 [PMID: 28625518] (WB, ICC/IF, Human)
• Katsura A, Tamura Y, Hokari S et al. ZEB1-regulated inflammatory phenotype in breast cancer cells. Mol Oncol. 2017 Jun 15 [PMID: 28618162] (WB, Human)
• Kim YH, Choi YW, Lee J et al. Senescent tumor cells lead the collective invasion in thyroid cancer Nat Commun. 2017 May 09 [PMID: 28489070] (IHC-P, Human)
• Osada A, Endo K, Kimura Y et al. Addiction of mesenchymal phenotypes on the FGF/FGFR axis in oral squamous cell carcinoma cells BioRxiv (WB, Human)
• Yoon C, Cho SJ, Chang KK et al. Role of Rac1 pathway in epithelial-to-mesenchymal transition and cancer stem-like cell phenotypes in gastric adenocarcinoma. Mol. Cancer Res. May 1 2017 [PMID: 28461325] (Human)
• Wang Y, Xu X, Yu S et al. Systematic characterization of A-to-I RNA editing hotspots in microRNAs across human cancers. Genome Res. Apr 14 2017 [PMID: 28411194] (Human)
• Sinh ND, Endo K, Miyazawa K, Saitoh M. Ets1 and ESE1 reciprocally regulate expression of ZEB1/ZEB2, dependent on ERK1/2 activity, in breast cancer cells. Cancer Sci. May 1 2017 [PMID: 28247944] (WB, Human)
• Guo E, Wang Z, Wang S. MiR-200c and miR-141 inhibit ZEB1 synergistically and suppress glioma cell growth and migration. Eur Rev Med Pharmacol Sci 2016 Aug [PMID: 27608897]
• Yu QC, Song W, Wang D, Zeng YA. Identification of blood vascular endothelial stem cells by the expression of protein C receptor Cell Res. 2016 Jul 01 [PMID: 27364685] (IHC)
• Muto Y, Suzuki K, Kato T et al. Heterogeneous expression of zinc-finger E-box-binding homeobox 1 plays a pivotal role in metastasis via regulation of miR-200c in epithelial-mesenchymal transition. Int J Oncol 2016 Sep [PMID: 27315529] (Human)
• Werden SJ, Sphyris N, Sarkar TR et al. Phosphorylation of serine 367 of FOXC2 by p38 regulates ZEB1 and breast cancer metastasis, without impacting primary tumor growth. Oncogene Jun 13 2016 [PMID: 27292262] (WB, Human)
• Sanfrancesco J, McKenney JK, Leivo MZ et al. Sarcomatoid Urothelial Carcinoma of the Bladder: Analysis of 28 Cases With Emphasis on Clinicopathologic Features and Markers of Epithelial-to-Mesenchymal Transition. Arch. Pathol. Lab. Med. 2016 Mar 31 [PMID: 27031776] (IHC-P, Human)
• Zhang L, Ma T, Brozick J et al. Effects of Kras activation and Pten deletion alone or in combination on MUC1 biology and epithelial-to-mesenchymal transition in ovarian cancer. Oncogene. 2016 Mar 14 [PMID: 26973247] (WB, Mouse)
• Incio J, Suboj P, Chin SM et al. Metformin Reduces Desmoplasia in Pancreatic Cancer by Reprogramming Stellate Cells and Tumor-Associated Macrophages. PLoS ONE. 2015 Dec 08 [PMID: 26641266] (WB, Human)
• Zheng X, Carstens JL, Kim J et al. Epithelial-to-mesenchymal transition is dispensable for metastasis but induces chemoresistance in pancreatic cancer. Nature. 2015 Nov 26 [PMID: 26560028] (IHC-P, Mouse)
• Yoon C, Cho SJ, Aksoy BA et al. Chemotherapy resistance in diffuse type gastric adenocarcinoma is mediated by RhoA activation in cancer stem-like cells Clin Cancer Res. 2015 Oct 18 [PMID: 26482039] (WB, Human)
• Ran J, Lin DL, Wu RF et al. ZEB1 promotes epithelial-mesenchymal transition in cervical cancer metastasis. Fertil Steril 2015 Jun [PMID: 25963537]
• Liu YN, Yin J, Barrett B et al. Loss of Androgen Regulated miR-1 Activates SRC and Promotes Prostate Cancer Bone Metastasis Mol. Cell. Biol. 2015 Mar 23 [PMID: 25802280] (WB, Mouse)
• Nolan KD, Franco OE, Hance MW et al. Tumor secreted Hsp90 subverts Polycomb function to drive prostate tumor growth and invasion J. Biol. Chem. 2015 Feb 10 [PMID: 25670862] (WB, Human)
• Sinigaglia A, Lavezzo E, Trevisan M et al. Changes in microRnA expression during disease progression in patients with chronic viral hepatitis. Liver Int. 2014 Nov 24 [PMID: 25417901] (WB)
• Fukagawa A, Ishii H, Miyazawa K, Saitoh M. Delta-EF1 associates with DnMT1 and maintains DnA methylation of the E-cadherin promoter in breast cancer cells. Cancer Med. 2014 Oct 15 [PMID: 25315069]
• Wang RT,Xu M,Song ZG et al. Decreased Expression of miR-216a Contributes to Non-small Cell Lung Cancer Progression Clin. Cancer Res. 2014 Jun 23 [PMID: 24958806] (WB, Human)
  
  Details:
  Zeb1 antibody used for WB in stable expression of miR216a or miR216a-antisense cells - A549 and H23 (Figure 4B), HepG2 cells (transfected with pre-oligo or pre-miR-216a plasmids, Supplementary Fig 6A) and A549 cells transfected with miR-216a, eIF4B and ZEB1 (Supplementary Fig 7).
• Yoshida T, Ozawa Y, Kimura T et al. Eribulin mesilate suppresses experimental metastasis of breast cancer cells by reversing phenotype from epithelial-mesenchymal transition (EMT) to mesenchymal-epithelial transition (MET) states. Br. J. Cancer 2014 Feb 26 [PMID: 24569463] (IHC-P, Human)
• Nam KH, Kim MA, Choe G et al. Deregulation of the cell polarity protein Lethal giant larvae 2 (Lgl2) correlates with gastric cancer progression. Gastric Cancer 2013 Dec 13 [PMID: 24337435] (IHC-P, Human)
• Gore AJ, Deitz SL, Palam LR et al. Pancreatic cancer-associated retinoblastoma 1 dysfunction enables TGF-beta to promote proliferation. J Clin Invest 2014 Jan 2 [PMID: 24334458] (IHC-P, ICC/IF, WB, Human, Mouse)
• Boyer JG, Murray LM, Scott K et al. Early onset muscle weakness and disruption of muscle proteins in mouse models of spinal muscular atrophy. Skelet Muscle. 2013 Oct 11 [PMID: 24119341] (WB, Mouse)
• Shi M, Han W, Spivack SD. A quantitative method to identify microRNAs targeting a messenger RNA using a 3'UTR RNA affinity technique. Anal Biochem. 2013 Aug 9 [PMID: 23938772] (WB, Human)
• Rajabi H, Alam M, Takahashi H et al. MUC1-C oncoprotein activates the ZEB1/miR-200c regulatory loop and epithelial-mesenchymal transition. Oncogene 2013 Apr 15 [PMID: 23584475] (ICC/IF, Human)
• Ding W, Dang H, You H et al. miR-200b restoration and DNA methyltransferase inhibitor block lung metastasis of mesenchymal-phenotype hepatocellular carcinoma. Oncogenesis 2012 [PMID: 23552699]
• Shin JO, Nakagawa E, Kim EJ, Cho KW, Lee JM, Cho SW, Jung HS. miR-200b regulates cell migration via Zeb family during mouse palate development. Histochem Cell Biol;137(4):459-70. 2012 Apr. [PMID: 22261924] (WB, IHC, Mouse)
• Saforo DJO Characterizing the role of extracellular matrix in non-small cell lung carcinoma metastasis using a 3D microenvironment mimetic in-vitro culture system. Thesis 2019 Aug (WB, Human)
• Ryu HS, Park do J, Kim HH, Kim WH, Lee HS. Combination of epithelial-mesenchymal transition and cancer stem cell-like phenotypes has independent prognostic value in gastric cancer. Hum Pathol;43(4):520-8. 2012 Apr. [PMID: 22018628] (IHC, Human)
• Kizuka Y, Kitazume S, Yoshida M, Taniguchi N Brain-specific expression of N-acetylglucosaminyltransferase IX (GnT-IX) is regulated by epigenetic histone modifications. J Biol Chem. [PMID: 21771782] (GS, Mouse)
• Gregory PA, Bracken CP, Smith E et al. An autocrine TGF-beta/ZEB/miR-200 signaling network regulates establishment and maintenance of epithelial-mesenchymal transition. Mol Biol Cell 2011 May [PMID: 21411626]

Reviews for ZEB1 Antibody (NBP1-05987) (6)

Average Rating: 4.8

(Based on 6 reviews)

We have 6 reviews tested in 2 species: Human, Mouse.

reviewed by: Anonymous WB Human 11/12/2017

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell lines
• Species: Human
• Lot: A1

Comments

• Comments: Sample: Whole cell lysates of of two cancer cell lines (10ug)
  Blocking: 5% nonfat dry milk reconstituted in TBS-T for 1 hour at room temperature
  Primary Antibody diluted (1:2000) in 5% nonfat dry milk reconstituted in TBS-T overnight at 4deg C
  Secondary Antibody: Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution in TBS-T

reviewed by: Melissa Skibba WB Mouse 05/23/2017

Summary

• Application: Western Blot
• Sample Tested: kidney,Mouse kidney
• Species: Mouse

Comments

• Comments: Performed on whole kidney in mouse with a 1:1000 primary and a 1:5000 secondary dilution.

reviewed by: Adrienne Franks IHC Mouse 04/18/2017

Summary

• Application: Immunohistochemistry
• Sample Tested: mouse liver,Mouse Pancreas,Mouse Spleen
• Species: Mouse

reviewed by: Anonymous WB Human 11/11/2011

Summary

• Application: Western Blot
• Sample Tested: Human
• Species: Human
• Comments: Great!

Blocking

• Blocking Details: Blocking Buffer: 5% Milk, Blocking Time: 1 hour

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:2000, Primary Ab Incubation Time: overnight

Secondary Antibody

• Secondary Description: Secondary Ab: Anti-mouse, Secondary Ab Dilution Ratio: 1:10000

Details

• Detection Notes: Detection Method: ECL

Comments

• Comments: Great!

reviewed by: Kelsey Stockbridge IHC-P Mouse 05/09/2011

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: liver and spleen
• Species: Mouse
• Lot: A1
• Comments: Nuclear expression in: liver (endothelial and mesenchymal cells) and spleen. Two different antigen retrieval and two dilutions were tested : EDTA and Citrate buffer, antibody dilution: 1:200-1:500- each with Biotin Streptavidin-HRP amplification. Best conditions with Citrate buffer, as antigen retrieval, antibody dilution at 1:500.

Blocking

• Blocking Details: Blocking Buffer: 10% FCS

Primary Anitbody

• Dilution Ratio: Dilution Ratio: 1:500, Incubation Dilution Buffer: 10% FCS in PBS 0.1% tween, Incubation Time: overnight, Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab: anti Rabbit-biotin then streptavidin-HRP, Secondary Ab Dilution: 1:200

Details

• Detection Notes: Detection Method: ABC system, Positive Control: liver and spleen, Antigen Retrieval: Citrate, 1 hour at 99 degrees
• Fixation Details: Sample Preparation: Formalin fixed

Comments

• Comments: Nuclear expression in: liver (endothelial and mesenchymal cells) and spleen. Two different antigen retrieval and two dilutions were tested : EDTA and Citrate buffer, antibody dilution: 1:200-1:500- each with Biotin Streptavidin-HRP amplification. Best conditions with Citrate buffer, as antigen retrieval, antibody dilution at 1:500.

reviewed by: Bhishma Amlani WB Mouse 05/28/2010

Summary

• Application: Western Blot
• Sample Tested: Murine Cell Line, Sample Amount: 50ug
• Species: Mouse
• Lot: A1
• Comments: Very clean antibody. However, endogenous and exogenous murine Zeb1 is detected at ~100 kDa.

Blocking

• Blocking Details: Blocking Buffer: 5% BSA in TBST, Blocking Time: 1 hour, Blocking Temp: 25 degrees Celsius

Primary Anitbody

• Dilution Ratio: Primary Ab Dilution Ratio: 1:2000, Incubation Dilution Buffer: 5% BSA in TBST, Incubation Time: 16hr, Incubation Temp: 4C

Secondary Antibody

• Secondary Description: Secondary Ab: HRP conjugated, donkey anti-rabbit, Secondary Ab Dilution Ratio: 1:10000
• Secondary Manufacturer Cat#: NA934V

Details

• Detection Notes: ECL, Exposure Time: 30sec, Ctrls: Ctrl Construct (lane 1), Overexpression Construct (lane 2), Specific Band: 100kDa

Comments

• Comments: Very clean antibody. However, endogenous and exogenous murine Zeb1 is detected at ~100 kDa.

FAQs for ZEB1 Antibody (NBP1-05987). (Showing 1 - 2 of 2 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.
2. We have an inquiry regarding the molecular weight of NBP1-05987, Anti-ZEB1. In the datasheet and customer review, the band sizes are very different (upper images: greater than 171kDa, under image (customer review): about 100kDa). If you have any idea why such difference occurred, could you let us know?
   • ZEB1 protein is 1,117 amino acids long and its expected molecular weight is 122.5 kDa. However, because it can undergo multiple types of potential post-translational modifications (including Isopeptide bond, Phosphoprotein, Ubl conjugation, see: Q64318), its observed molecular weight is always different than the predicted one. In our validation studies we observed specific bands in human and mouse cells at approximately 171kDa and almost all of the commercially available antibodies detect the target bands in this range (or at approximately 200kD). This product has been cited in the Journal of Biological Chemistry by Kizuka et al 2011 and they found our antibody to detect the target band at approximately 170kD (Please refer to Supplementary Figure # 3; JBC September 9, 2011 vol. 286 no. 36 31875-31884).

Bioinformatics

• Gene Symbol: ZEB1
• Entrez:
  • 6935(Human)
  • 21417(Mouse)
• Uniprot:
  • P37275()