Anti-Nanog antibody (ab80892)

Rabbit polyclonal Nanog antibody. Validated in WB, IHC, ICC, Flow Cyt, ICC/IF and tested in Mouse. Cited in 179 publication(s). Independently reviewed in 18 review(s).

Overview

  • Product name
  • Description
    Rabbit polyclonal to Nanog
  • Host species
    Rabbit
  • Tested applications
    Suitable for: Flow Cyt, IHC - Wholemount, ICC, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Recombinant full length protein corresponding to Mouse Nanog.

  • Positive control
    • ICC/IF: Mouse embryonic stem cells. WB: Mouse embryonic stem cell line D3. IHC-P: Mouse teratoma tissue. IHC-Wm: Mouse pre-implantation embryo. Flow Cyt: Mouse embryonic stem cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab80892 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
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

IHC - Wholemount Use at an assay dependent concentration.
ICC 1/150 - 1/700.
WB 1/300 - 1/2000. Detects a band of approximately 35 kDa (predicted molecular weight: 35 kDa).
IHC-P 1/400. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    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.
  • Tissue specificity
    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.
  • Sequence similarities
    Belongs to the Nanog homeobox family.
    Contains 1 homeobox DNA-binding domain.
  • Developmental stage
    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.
  • Cellular localization
    Nucleus.
  • 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

Images

  • Spatio-temporal changes in expression of pluripotency markers and self-organization of stem cell-derived aggregates in 3D culture.
    Panels A-D:
    Expression of the pluripotency marker Nanog decreases as vesicles mature between differentiation days 5–8.

    Panels A’-D’: Progression of outer epithelium ruffling indicating self-organization.

    Three mouse embryonic stem cell aggregates were fixed with 4% paraformaldehyde, then cryoprotected with a serial treatment of 15% and 30% sucrose and embedded in tissue freezing medium. Frozen tissue blocks were sectioned into 10 or 12 μm cyrosections. For immunostaining, a 3% goat or horse serum and 0.1% Triton-X100 solution was used for primary antibody incubation. An Alexa Fluor® 488, 568, or 647 conjugated anti-mouse IgG or anti-goat IgG and an Alexa Fluor 568 or 647 conjugated anti-rabbit IgG were used as secondary antibodies. A DAPI counterstain was used to visualize cellular nuclei.

    Microscopy was performed on a Nikon TE2000 Inverted Microscope or an Olympus FV1000-MPE Confocal/Multiphoton Microscope.

  • ab80892 staining Nanog in mouse embryonic stem cells by immunocytochemistry/ immunofluorescence.

    Cells were paraformaldehyde fixed and permeabilized in Tween-20 prior to blocking in 20% serum for 30 minutes at 25°C. The primary antibody was diluted 1/100 and incubated with the sample for 4 hour at 25°C. A Cy®5 conjugated goat polyclonal to rabbit IgG antibody, diluted 1/100, was used as the secondary.

    Nuclear staining with DAPI.

    See Abreview

  • All lanes : Anti-Nanog antibody (ab80892) at 1/500 dilution

    Lane 1 : Marker
    Lanes 2-3 : Mouse embryonic stem cell line D3
    Lanes 4-5 : Mutant mouse embryonic stem cell line known to not express nanog

    Lysates/proteins at 200000 cells per lane.

    Secondary
    All lanes : goat anti-rabbit HRP at 1/2000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 35 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?


    Exposure time: 10 minutes


    Blocked with milk for 30 minutes.

    See Abreview

  • ab80892 staining mouse teratoma sections by IHC-P. The tissue was fixed with formaldehyde and a heat mediated antigen retrival step was performed with citric acid pH 6. Blocking of the sample was done with 1% BSA for 10 minutes at 21°C, followed by staining with ab80892 at 1/400 in TBS/BSA/azide for 2h at 21°C. A biotinylated goat anti-rabbit polyclonal antibody at 1/200 was used as the secondary antibody.

    See Abreview

  • IHC-Wholemount image of anti-Nanog antibody (ab80892) staining on a mouse pre-implantation embryo.

    This embryo was fixed in paraformaldehyde, permeabilized in 0.25% Triton X and then blocked in 5% serum for 1 hour at room temperature. It was stained with anti-Nanog antibody overnight at 4 degrees (right), and DAPI (left).

    See Abreview

  • Overlay histogram of Nanog staining in mouse embryonic stem cells using ab80892, at 1:100 dilution.

    Purple histogram represents negative control (rabbit IgG).

    Green line represents anti-Nanog antibody (ab80892).

    See Abreview

References

This product has been referenced in:
  • Wang Y  et al. Generation of induced pluripotent stem cell line (ZZUi0013-A) from a 65-year-old patient with a novel MEOX2 gene mutation in Alzheimer's disease. Stem Cell Res 34:101366 (2019). Read more (PubMed: 30616143) »
  • Wickremesekera AC  et al. Expression of cancer stem cell markers in metastatic melanoma to the brain. J Clin Neurosci 60:112-116 (2019). Read more (PubMed: 30626524) »
See all 200 Publications for this product

Customer reviews and Q&As

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1-10 of 18 Abreviews

Application
Immunocytochemistry
Sample
Rabbit Cultured Cells (brain)
Permeabilization
Yes - tritonx100
Specification
brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde

Kyungjoo Seong

Verified customer

Submitted Oct 31 2016

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lung)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6.0
Permeabilization
No
Specification
lung
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1.5% · Temperature: 25°C
Fixative
10% Formalin

Abcam user community

Verified customer

Submitted Apr 25 2016

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Embryonic)
Permeabilization
Yes - Triton-X100 (0.5%)
Specification
Embryonic
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Oct 23 2015

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Tissue lysate - whole (mouse cerebellum)
Specification
mouse cerebellum
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Nov 18 2014

Application
Flow Cytometry
Fixation
Paraformaldehyde
Permeabilization
Yes - 0.5% v/v tween20 in PBS
Sample
Mouse Cell (mouse embryonic stem cell)
Specification
mouse embryonic stem cell
Gating Strategy
live cells
Preparation
Cell harvesting/tissue preparation method: cell dissociation buffer
Sample buffer: 0.5% v/v tween20 in PBS

Abcam user community

Verified customer

Submitted Apr 28 2014

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (lung)
Specification
lung
Fixative
Formaldehyde
Permeabilization
Yes - with Tris and Tween
Blocking step
Peroxidase as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: 24°C

Abcam user community

Verified customer

Submitted May 15 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: TRIS-EDTA Buffer pH 9,0
Permeabilization
Yes - Wash Buffer from Dako with Tween
Specification
Tonsil
Fixative
Paraformaldehyde

Mr. Rudolf Jung

Verified customer

Submitted Mar 22 2013

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Common marmoset Tissue sections (embryonic testis E92)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako antigen retrieval solution
Permeabilization
No
Specification
embryonic testis E92
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Dr. 煜欽 林

Verified customer

Submitted Feb 11 2013

Application
Western blot
Loading amount
200000 cells
Gel Running Conditions
Reduced Denaturing (10% gel)
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cell)
Specification
mouse embryonic stem cell
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 16°C

Abcam user community

Verified customer

Submitted Oct 24 2012

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Teratoma)
Specification
Teratoma
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 21°C

Mr. Carl Hobbs

Verified customer

Submitted Apr 12 2012

1-10 of 18 Abreviews

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