Overview

  • Product nameAnti-Nanog antibody - ChIP GradeSee all Nanog primary antibodies ...
  • Description
    Rabbit polyclonal to Nanog - ChIP Grade
  • Tested applicationsIHC-P, ICC, Flow Cyt, ICC/IF, WB, Sandwich ELISA, ChIPmore details
  • Species reactivity
    Reacts with: Human

    Does not react with

    Mouse
  • Immunogen

    Synthetic peptide:

    CGPEENYPSLQMSSAEMPHTE

    , corresponding to amino acids 29-49 of Human Nanog.

Properties

Applications

Our Abpromise guarantee covers the use of ab21624 in the following tested applications.

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

Application Abreviews Notes
IHC-P Use at an assay dependent concentration.
ICC 1/100.
Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/1000.
WB 1/200. Detects a band of approximately 38 kDa (predicted molecular weight: 34 kDa).

Some users report a stronger signal after blocking and diluting antibodies with 1% BSA instead of 5% milk.

Sandwich ELISA Use a concentration of 0.5 µg/ml. Can be paired for Sandwich ELISA with Mouse monoclonal [1E6C4] to Nanog (ab76586). For sandwich ELISA, use this antibody as Detection at 0.5 µg/ml with Mouse monoclonal [1E6C4] to Nanog (ab76586) as Capture.
ChIP Use at an assay dependent concentration. PubMed: 18710938

Target

  • FunctionTranscription 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.
  • Tissue specificityExpressed 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.
  • Sequence similaritiesBelongs to the Nanog homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Developmental stageExpressed 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.
  • Cellular localizationNucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 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 - ChIP Grade images

  • All lanes : Anti-Nanog antibody - ChIP Grade (ab21624) at 1/200 dilution

    Lane 1 : Human fetal fibroblast lysate
    Lane 2 : Human iPS cell lysate


    Predicted band size : 34 kDa
  • ab21624 staining Nanog in Human H9 differentiated stem cells by Flow Cytometry. The sample was incubated with the primary antibody (1/100 in PBS + 5% FBS and 1% saponin) for 1 hour at 4°C. A FITC-conjugated Human anti-rabbit IgG polyclonal (1/400) was used as the secondary antibody.

    See Abreview

  • Standard Curve for Nanog (Analyte: Nanog protein (ab50053)); dilution range 1 pg/ml to 1 ug/ml using Capture Antibody Mouse monoclonal [1E6C4] to Nanog (ab76586) at 0.2 ug/ml and Detector Antibody Rabbit polyclonal to Nanog - ChIP Grade (ab21624) at 0.5 ug/ml.
  • ab21624 at a 1/1000 dilution staining Nanog in human embryonic stem cells by ICC. An antibody towards Oct4, which has an identical staining pattern to ab21624, was used as a positive control.ab21624 at a 1/1000 dilution staining Nanog in human embryonic stem cells by ICC. An antibody towards Oct4, which has an identical staining pattern to ab21624, was used as a positive control.
  • All lanes : Anti-Nanog antibody - ChIP Grade (ab21624) at 1/1000 dilution

    Lane 1 : whole cell extract of mouse 3T3 cells expressing mouse Nanog.
    Lane 2 : whole cell extract of mouse 3T3 cells expressing human Nanog.


    Predicted band size : 34 kDa
    Observed band size : 38 kDa (why is the actual band size different from the predicted?)
    ab21624 recognizes human Nanog but does not recognize mouse Nanog in Western Blot.
  • ab21624 staining Nanog in Human testis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 20°C; antigen retrieval was by heat mediation with a citrate buffer (pH6). Samples were incubated with primary antibody (1/500 in PBS) for 12 hours at 4°C. A Biotin-conjugated Goat anti-rabbit polyclonal (1/200) was used as the secondary antibody.

    See Abreview

Protocols

References for Anti-Nanog antibody - ChIP Grade (ab21624)

This product has been referenced in:
  • Nie Y  et al. Scalable passaging of adherent human pluripotent stem cells. PLoS One 9:e88012 (2014). ICC/IF ; Human . Read more (PubMed: 24498239) »
  • Liu B  et al. Nanog1 in NTERA-2 and recombinant NanogP8 from somatic cancer cells adopt multiple protein conformations and migrate at multiple M.W species. PLoS One 9:e90615 (2014). WB ; Human . Read more (PubMed: 24598770) »

See all 23 Publications for this product

Product Wall

Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - Permeabilization Buffer
Sample Human Cell (Fanconi's Anaemia iPSC)
Specification Fanconi's Anaemia iPSC
Gating Strategy Isotype negative control (white)
Preparation Cell harvesting/tissue preparation method: Typsin-EDTA Cell Dissociation
Sample buffer: PBS and 10% FBS
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Verified customer

Submitted Jan 29 2014

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Application Flow Cytometry
Fixation Paraformaldehyde
Permeabilization Yes - 0.5% Saponin Buffer
Sample Human Cell (H9 Human Embryonic Stem Cells)
Specification H9 Human Embryonic Stem Cells
Gating Strategy FL1/FSC on isotype control
Preparation Cell harvesting/tissue preparation method: Single cell dissociation buffer
Sample buffer: FACS Buffer (5% FBS)
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Submitted Jul 19 2013

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Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 37°C
Sample Human Cell (Embryonic Stem Cell)
Specification Embryonic Stem Cell
Permeabilization Yes - BD Perm/Wash Buffer
Fixative Paraformaldehyde
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Submitted May 22 2013

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Application Flow Cytometry
Sample Human Cell (Embryonic Stem Cell)
Specification Embryonic Stem Cell
Preparation Cell harvesting/tissue preparation method: Trypsinisation
Sample buffer: BD Perm/Wash Buffer
Fixation Paraformaldehyde
Permeabilization Yes - BD Perm/Wash Buffer
Gating Strategy Gating position dependent on isotype control population
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Submitted May 08 2013

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (liver)
Specification liver
Fixative Paraformaldehyde
Permeabilization Yes - triton-100
Blocking step Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
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Submitted Dec 10 2012

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Mouse ES cells exogenously expressing Human Nanog)
Specification Mouse ES cells exogenously expressing Human Nanog
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% triton-X
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 16°C
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Submitted Oct 22 2012

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Application ChIP
Sample Human Cell lysate - whole cell (liver cancer)
Specification liver cancer
Type Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: Paraformaldehyde
Detection step Other
Positive control Input
Negative control IgG
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Submitted Oct 18 2012

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Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (liver cancer)
Specification liver cancer
Fixative Paraformaldehyde
Permeabilization Yes - Triton-100
Blocking step Serum as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
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Submitted Oct 16 2012

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Application Western blot
Sample Human Cell lysate - whole cell (liver cancer)
Loading amount 10 µg
Specification liver cancer
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Submitted Oct 11 2012

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab34222 and one vial of ab13537.

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