Nanog Antibody - BSA Free

Western Blot: Nanog Antibody [NB100-588] - ZNF217 and ALKBH5 regulate NANOG and KLF4 expression via modulation of m6A levels. MCF-7 subclones were exposed to 20% or 1% O2 for 48 h and immunoblot assays were performed. H. MCF-7 subclones were exposed to 20% or 1% O2 for 48 h. The percentage of m6A+ KLF4 mRNA was determined. *P < 0.05, **P < 0.01 vs. NTC at 20% O2; ##P < 0.01 vs. NTC at 1% O2 by two-way ANOVA. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/fulltext/11743), licensed under a CC-BY license.

Flow Cytometry: Nanog Antibody [NB100-588] - FACS analysis of Nanog expression in primary mouse hepatic stellate cell cultured for 7 days. Image from verified customer review.

Western Blot: Nanog Antibody [NB100-588] - Detection of Mouse Nanog by Western Blot and Immunoprecipitation.

Western Blot: Nanog Antibody [NB100-588] - The expression of pluripotency factor Nanog in human head and neck cancer cell lines cell lines FaDu and HN5. WB from a verified customer review.

Western Blot: Nanog Antibody [NB100-588] - Detection of mouse Nanog by western blot. Samples: Whole cell lysate (50 ug) from F9 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-Nanog antibody NB100-588 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.

Western Blot: Nanog Antibody [NB100-588] - HIFs are required for hypoxia-induced expression of pluripotency factors. ZR75.1 cells were treated with vehicle or digoxin (200 nM), exposed to 20% or 1% O2 for 48 h, and HIF-1alpha, NANOG and SOX2 immunoblot assays were performed. Image collected and cropped by CiteAb from the following publication (//www.oncotarget.com/fulltext/11743), licensed under a CC-BY license.

Flow Cytometry: Nanog Antibody [NB100-588] - staining of NTERA-2 cells using NB100-588 at a 1:50 dilution detected using Dylight-488 conjugated goat anti-rabbit secondary antibody.

Western Blot: Nanog Antibody [NB100-588] - HIFs are required for hypoxia-induced expression of pluripotency factorsA-C. Breast cancer cell lines were exposed to 20% or 1% O2 for 24 h & NANOG (A), KLF4 (B), & SOX2 (C) mRNA levels were determined by RT-qPCR, relative to 18S rRNA, & normalized to the mean value for MDA-MB-231 cells (MDA231) at 20% O2 (mean ± SEM; n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 vs. same cell line at 20% O2 by Student's t test. D & E. HCC-1954 (D) & MCF-7 (E) subclones, which were stably transfected with an expression vector encoding a non-targeting control (NTC) shRNA, or vector encoding shRNA targeting HIF-1 alpha (sh1 alpha ) or HIF-2 alpha (sh2 alpha ), or vectors encoding shRNAs targeting both HIF-1 alpha & HIF-2 alpha (DKD), were exposed to 20% or 1% O2 for 24 h & RT-qPCR was performed to determine NANOG (D) or KLF4 (E) mRNA levels relative to 18S rRNA. The results were normalized to NTC at 20% O2 (mean ± SEM; n = 3). *P < 0.05, **P < 0.01, ***P < 0.001 vs. NTC at 20% O2; #P < 0.05, ##P < 0.01, ###P < 0.001 vs. NTC at 1% O2 by ANOVA. F. ZR75.1 cells treated with vehicle or digoxin (200 nM) were exposed to 20% or 1% O2 for 24 h & SOX2 mRNA was measured (mean ± SEM; n = 3). *P < 0.05, **P < 0.01 vs. NTC at 20% O2; ###P < 0.001 vs. NTC at 1% O2 by ANOVA. G & H. NTC & DKD subclones of HCC-1954 (G) & MCF-7 (H) were exposed to 20% or 1% O2 for 48 h, whole cell lysates were prepared, & immunoblot assays were performed to analyze HIF-1 alpha , HIF-2 alpha , NANOG & KLF4 protein expression. Actin was also analyzed as a loading control. I. ZR75.1 cells were treated with vehicle or digoxin (200 nM), exposed to 20% or 1% O2 for 48 h, & HIF-1 alpha , NANOG & SOX2 immunoblot assays were performed. Image collected & cropped by CiteAb from the following publication (//www.oncotarget.com/lookup/doi/10.18632/oncotarget.11743), licensed under a CC-BY license. Not internally tested by Novus Biologicals.

• Reactivity: Hu, Mu
• Applications: WB, Flow, ICC/IF, IP
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

Nanog Antibody - BSA Free Summary

• Immunogen: A synthetic peptide which maps to a region between residues 250 and the C-terminus (residue 305) of mouse Nanog homeobox using the numbering given in TrEMBL entry Q80Z64 (GeneID 71950).
• Marker: Embryonic Stem Cell Marker
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: NANOG
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Flow Cytometry 1:150
  • Immunocytochemistry/ Immunofluorescence 1:10-1:2000
  • Immunoprecipitation 2-5 ug/mg lysate
  • Western Blot 1:2000-1:10000
• Application Notes: WB from a verified customer review, PMID ( 28228260). ICC/IF reported in (PMID: 22505032). Nanog antibody validated for FLOW from a verified customer review.

Reactivity Notes

Human reactivity reported in (PMID: 28228260), Customer review.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.
• Buffer: Tris-Citrate/Phosphate (pH 7.0 - 8.0)
• Preservative: 0.09% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for Nanog Antibody - BSA Free

• FLJ12581
• FLJ40451
• hNanog
• homeobox protein NANOG
• Homeobox transcription factor Nanog
• homeobox transcription factor Nanog-delta 48
• Nanog homeobox
• Nanog

Background

Nanog is one of the molecular markers suitable for recognizing the undifferentiated state of stem cells in the mouse and human. It is a divergent homeodomain protein that directs propagation of undifferentiated ES cells. Nanog mRNA is present in pluripotent mouse and human cell lines, and absent from differentiated cells. Nanog-deficient ES cells lose pluripotency and differentiate into extraembryonic endoderm lineage.

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 Nanog Antibody (NB100-588)(15)

Publications using NB100-588

• P Montero, J Milara, M Pérez-Leal, C Estornut, I Roger, A Pérez-Fida, C Sanz, J Cortijo Paclitaxel-Induced Epidermal Alterations: An In Vitro Preclinical Assessment in Primary Keratinocytes and in a 3D Epidermis Model International Journal of Molecular Sciences, 2022-01-20;23(3):. 2022-01-20 [PMID: 35163066]
• Nanduri J, Wang N, Wang Bl, Prabhakar Nr Lysine demethylase KDM6B regulates HIF-1 alpha mediated systemic and cellular responses to intermittent hypoxia Physiological genomics 2021-07-23 [PMID: 34297635]
• Zhang C, Huang S, Zhuang H et al. YTHDF2 promotes the liver cancer stem cell phenotype and cancer metastasis by regulating OCT4 expression via m6A RNA methylation Oncogene 2020-05-04 [PMID: 32366907]
• Daum R, Brauchle EM, Berrio DAC et al. Non-invasive detection of DNA methylation states in carcinoma and pluripotent stem cells using Raman microspectroscopy and imaging Sci Rep 2019-05-07 [PMID: 31065074] (ICC/IF, Human, Mouse)
• Lu H, Chen I, Shimoda LA et al. Chemotherapy-Induced Ca 2+ Release Stimulates Breast Cancer Stem Cell Enrichment Cell Rep. 2017-02-21 [PMID: 28228260] (WB, Human)
  
  Details:
  The positive impact of HIF inhibitors on breast cancer chemotherapy is explored through GSTO1 knockdown.
• Zhang C, Zhi WI, Lu H et al. Hypoxia-inducible factors regulate pluripotency factor expression by ZNF217- and ALKBH5-mediated modulation of RNA methylation in breast cancer cells. Oncotarget. 2016-08-31 [PMID: 27590511] (WB, Human)
• Zhang C, Samanta D, Lu H et al. Hypoxia induces the breast cancer stem cell phenotype by HIF-dependent and ALKBH5-mediated m6A-demethylation of NANOG mRNA. Proc Natl Acad Sci U S A. 2016-03-21 [PMID: 27001847] (WB, Human)
• Lu H Samanta D Xiang L et al. Chemotherapy triggers HIF-1-dependent glutathione synthesis and copper chelation that induces the breast cancer stem cell phenotype. Proc Natl Acad Sci U S A. 2015-07-30 [PMID: 26229077] (WB, Human)
• Kohler Erin E, Baruah Jugajyoti, Urao Norifumi et al. Low-dose 6-bromoindirubin-3'-oxime induces partial dedifferentiation of endothelial cells to promote increased neovascularization. Stem Cells 2014-01-01 [PMID: 24496925] (WB, Mouse)
• Hamada M, Malureanu LA, Wijshake T et al. Reprogramming to pluripotency can conceal somatic cell chromosomal instability. PLoS Genet 2012-01-01 [PMID: 22952451]
• Wu Y, Guo Z, Wu H et al. SUMOylation represses Nanog expression via modulating transcription factors Oct4 and Sox2. PLoS One 2012-01-01 [PMID: 22745796]
• Ganuza M, Saiz-Ladera C, Canamero M, Gomez G, Schneider R, Blasco MA, Pisano D, Paramio JM, Santamaria D, Barbacid M. Genetic inactivation of Cdk7 leads to cell cycle arrest and induces premature aging due to adult stem cell exhaustion. EMBO J. 2012-04-13 [PMID: 22505032] (ICC/IF, Mouse)
• Morgado-Palacin L, Llanos S, Serrano M Ribosomal stress induces L11- and p53-dependent apoptosis in mouse pluripotent stem cells. Cell Cycle. 2012-02-01 [PMID: 22262176] (ICC/IF, Mouse)
• Kohler EE, Cowan CE, Chatterjee I et al. NANOG induction of fetal liver kinase-1 (FLK1) transcription regulates endothelial cell proliferation and angiogenesis. Blood 2010-11-30 [PMID: 21119109] (WB, Human)
• Carey BW, Markoulaki S, Hanna J et al. Reprogramming of murine and human somatic cells using a single polycistronic vector. Proc Natl Acad Sci U S A 2009-01-01 [PMID: 19109433]

Reviews for Nanog Antibody (NB100-588) (4)

Average Rating: 3.8

(Based on 4 reviews)

We have 4 reviews tested in 2 species: Human, Mouse.

reviewed by: Verified Customer WB Human 10/11/2018

Summary

• Application: Western Blot
• Sample Tested: Human head and neck cancer cell lines
• Species: Human
• Lot: A-2

reviewed by: Verified Customer WB Human 08/22/2015

Summary

• Application: Western Blot
• Sample Tested: Human cancer cell whole cell lysate
• Species: Human

reviewed by: Verified Customer Flow Mouse 04/29/2014

Summary

• Application: Flow Cytometry
• Species: Mouse

reviewed by: Verified Customer WB Human 06/08/2011

Summary

• Application: Western Blot
• Sample Tested: HUVEC, MCF7, JURKAT, Sample Amount: 2-30 ug
• Species: Human

Bioinformatics

• Gene Symbol: NANOG
• Entrez:
  • 71950(Mouse)
• Uniprot:
  • Q80Z64()