RabMAb

Anti-Nanog抗体[EPR2027(2)] (ab109250)

概述

  • 产品名称Anti-Nanog抗体[EPR2027(2)]
    参阅全部 Nanog 一抗
  • 描述
    兔单克隆抗体[EPR2027(2)] to Nanog
  • 特异性100% identities with NANOGP8
  • 经测试应用适用于: WB, IHC-P, ICC/IF, Flow Cytmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human Nanog aa 1-100 (N terminal).
    Database link: Q9H9S0

  • 阳性对照
    • WB: NCCIT cell lysate. IHC-P: Human seminoma tissue, Human dysgerminoma tissue and Human embryonal carcinoma tissue. ICC/IF: Human embryonic carcinoma and Human liver cell lines.
  • 常规说明

    This product is a recombinant rabbit monoclonal antibody.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated ‘PUR’ on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    Alternative versions available:
    Anti-Nanog antibody (Alexa Fluor® 488) [EPR2027(2)] (ab196155)
    Anti-Nanog antibody (Alexa Fluor® 647) [EPR2027(2)] (ab195018)

性能

应用

Our Abpromise guarantee covers the use of ab109250 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB 1/1000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).
IHC-P 1/100 - 1/250.

See protocols (link: http://www.abcam.com/protocols/ihc-antigen-retrieval-protocol).

Antigen retrieval is recommended.

ICC/IF 1/100 - 1/250.
Flow Cyt 1/70.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

靶标

  • 功能Transcription regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes (By similarity). Acts as a transcriptional activator or repressor (By similarity). Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3' (By similarity). When overexpressed, promotes cells to enter into S phase and proliferation.
  • 组织特异性Expressed in testicular carcinoma and derived germ cell tumors (at protein level). Expressed in fetal gonads, ovary and testis. Also expressed in ovary teratocarcinoma cell line and testicular embryonic carcinoma. Not expressed in many somatic organs and oocytes.
  • 序列相似性Belongs to the Nanog homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • 发展阶段Expressed in embryonic stem (ES) and carcinoma (EC) cells. Expressed in inner cell mass (ICM) of the blastocyst and gonocytes between 14 and 19 weeks of gestation (at protein level). Not expressed in oocytes, unfertilized oocytes, 2-16 cell embryos and early morula (at protein level). Expressed in embryonic stem cells (ES). Expression decreases with ES differentiation.
  • 细胞定位Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • Embryonic stem cell specific homeobox protein (Nanog) antibody
    • ENK antibody
    • FLJ12581 antibody
    • hNanog antibody
    • Homeobox protein NANOG antibody
    • Homeobox transcription factor Nanog antibody
    • homeobox transcription factor Nanog-delta 48 antibody
    • NANOG antibody
    • Nanog homeobox antibody
    • NANOG_HUMAN antibody
    see all

Anti-Nanog antibody [EPR2027(2)] 图像

  • Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/10000 dilution (purified) + NCCIT cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size : 35 kDa
  • Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/1000 dilution (unpurified) + NCCIT cell lysate at 10 µg

    Predicted band size : 35 kDa
    Observed band size : 37 kDa (why is the actual band size different from the predicted?)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human seminoma tissue labelling Nanog with purified ab109250 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human seminoma tissue labelling Nanog with unpurified ab109250 at 1/100.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human dysgerminoma tissue labelling Nanog with unpurified ab109250 at 1/100.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human embryonal carcinoma tissue labelling Nanog with unpurified ab109250 at 1/100.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human normal colon tissue shows negative staining of Nanog with unpurified ab109250.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human adult kidney tissue shows negative staining of Nanog with unpurified ab109250.

  • Flow Cytometry analysis of NCCIT cells labelling Nanog with purified ab109250 at 1/70 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • Immunocytochemistry/Immunofluorescence analysis of NCCIT(human pluripotent embryonal carcinoma) cells labelling Nanog with purified ab109250 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

  • Immunocytochemistry/Immunofluorescence analysis of Human Liver cells labelling Nanog with unpurified ab109250. Cells were fixed with Paraformaldehyde, permeabilized with Triton X-100 0.1% and blocked with 1% BSA for 12 hours at 4°C. Sample was incubated with primary antibody (1/500 in PBS) for 16 hour at 4°C. An Alexa Fluor®647-conjugated Donkey anti-rabbit(1/1000) IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Immunocytochemistry/Immunofluorescence analysis of embryonic carcinoma cells labelling Nanog with unpurified ab109250 at 1/100.

Anti-Nanog antibody [EPR2027(2)] (ab109250)参考文献

This product has been referenced in:
  • Yang Y  et al. Heightened potency of human pluripotent stem cell lines created by transient BMP4 exposure. Proc Natl Acad Sci U S A 112:E2337-46 (2015). Read more (PubMed: 25870291) »
  • Lee K  et al. Engraftment of human iPS cells and allogeneic porcine cells into pigs with inactivated RAG2 and accompanying severe combined immunodeficiency. Proc Natl Acad Sci U S A 111:7260-5 (2014). ICC/IF ; Human . Read more (PubMed: 24799706) »

See all 7 Publications for this product

Product Wall

Application Western blot
Sample Human Cell lysate - whole cell (patient derived iPSC)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification patient derived iPSC
Blocking step Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 05 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Horse Cell (testis tissues)
Specification testis tissues
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative Paraformaldehyde
Username

Mr. Heejun Jung

Verified customer

提交于 Aug 19 2015

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (iPSC)
Permeabilization Yes - 0.25% TritonX
Specification iPSC
Blocking step BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 3%
Fixative Formaldehyde
Username

Abcam user community

Verified customer

提交于 Jul 22 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (seminoma tumor)
Specification seminoma tumor
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Leica Bond Epitope Retrieval 2
Permeabilization No
Username

Abcam user community

Verified customer

提交于 Apr 02 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Marmoset (common) Tissue sections (embryonic testis E92)
Specification embryonic testis E92
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
Permeabilization No
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

煜欽 林

Verified customer

提交于 Feb 08 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (SW480 cells, HCT116 cells)
Loading amount 25 µg
Specification SW480 cells, HCT116 cells
Gel Running Conditions Reduced Denaturing (4-12%)
Username

Abcam user community

Verified customer

提交于 Jan 17 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (liver cancer)
Loading amount 20 µg
Specification liver cancer
Gel Running Conditions Reduced Denaturing (10%gel)
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Oct 18 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (liver cancer)
Specification liver cancer
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: EDTA
Permeabilization Yes - Triton-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Oct 15 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (liver cancer)
Specification liver cancer
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton-100
Blocking step BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Username

Abcam user community

Verified customer

提交于 Oct 09 2012

Thank you for your telephone call this morning. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody. As discussed on the telephone, I would ilke to reassure you that ab109250 is covered by our 6 monthAbpromise g...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"