Anti-Nanog antibody (ab14959)

Reviews (2)Q&A (4)References (14)

Key features and details

  • Rabbit polyclonal to Nanog
  • Suitable for: WB, IHC-P
  • Reacts with: Mouse
  • Isotype: IgG

Overview

  • Product name

    Anti-Nanog antibody
    See all Nanog primary antibodies

  • Description

    Rabbit polyclonal to Nanog

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC/IF

  • Species reactivity

    Reacts with: Mouse
    Does not react with: Human

  • Immunogen

    Synthetic peptide corresponding to Mouse Nanog aa 250 to the C-terminus (C terminal) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab14960)

  • Positive control

    • This antibody gave a positive signal in the following whole cell lysates: Mouse Embryonic Stem Cell.

Properties

Applications

Our Abpromise guarantee covers the use of ab14959 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 34 kDa (predicted molecular weight: 34 kDa).
IHC-P 1/600. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC/IF.

    • 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

      • Western blot - Anti-Nanog antibody (ab14959)
        Western blot - Anti-Nanog antibody (ab14959)
        All lanes : Anti-Nanog antibody (ab14959) at 1 µg/ml

        Lanes 3-4 : Mouse Nanog peptide (ab14960) at 1 µg/ml

        Predicted band size: 34 kDa
        Observed band size: 34.6 kDa
        why is the actual band size different from the predicted?
        Additional bands at: 18 kDa (possible cross reactivity), 38 kDa (possible cross reactivity), 50 kDa (possible cross reactivity)



        Lanes 1-4 : Nanog antibody (ab14959) at 1 ug/ml
        Lanes 3-4 : Nanog peptide (ab14960) at 1 ug/ml

        Observed band size: 34.2 kDa
        Additional bands at: 18 kDa (possible cross reactivity), 38 kDa (possible cross reactivity), 50 kDa (possible cross reactivity).

        Lanes 1 and 3: Mouse embryonic stem cell lysate (positive control)
        Lanes 2 and 4: Human embryonic stem cell lysate (negative control)

        ab14959 recognised a band of the expected size for Nanog in mouse embryonic stem cells but not in human embryonic stem cells. Additional bands were also recognised in mouse embyonic stem cells.

        Performed under reducing conditions.

      • Western blot - Anti-Nanog antibody (ab14959)
        Western blot - Anti-Nanog antibody (ab14959)This image is courtesy of Adam Yates and Ian Chambers, University of Edinburgh
        All lanes : Anti-Nanog antibody (ab14959) at 0.8 µg/ml

        All lanes : ES Cell Nuclear Extract

        Predicted band size: 34 kDa

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody (ab14959)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody (ab14959)This image is courtesy of an Abreview submitted by Carl Hobbs

        ab14959 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 ab14959 at 1/600 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

      References

      Publishing research using ab14959? Please let us know so that we can cite the reference in this datasheet.

      ab14959 has been referenced in 14 publications.

      • Dong H  et al. GLI1 activation by non-classical pathway integrin avß3/ERK1/2 maintains stem cell-like phenotype of multicellular aggregates in gastric cancer peritoneal metastasis. Cell Death Dis 10:574 (2019). WB ; Human . PubMed: 31366904
      • Kim HJ  et al. Cyclin-dependent kinase 1 activity coordinates the chromatin associated state of Oct4 during cell cycle in embryonic stem cells. Nucleic Acids Res 46:6544-6560 (2018). PubMed: 29901724
      • Kwak S  et al. Zinc finger proteins orchestrate active gene silencing during embryonic stem cell differentiation. Nucleic Acids Res 46:6592-6607 (2018). WB ; Mouse . PubMed: 29846698
      • Kim TW  et al. Ctbp2-mediated ß-catenin regulation is required for exit from pluripotency. Exp Mol Med 49:e385 (2017). PubMed: 29026198
      • Shin J  et al. Aurkb/PP1-mediated resetting of Oct4 during the cell cycle determines the identity of embryonic stem cells. Elife 5:e10877 (2016). PubMed: 26880562
      View all Publications for this product

      Customer reviews and Q&As

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      1-6 of 6 Abreviews or Q&A

      Question

      Dear,

      I am afraid that you may missunderstand my comments. We have a clear positivity using this antibody. Our question is its sensitivity because of the reason I mentioned below.

      Looking forward to hearing from you soon.

      Best Wishes,

      Read More
      Answer

      Thank you for your response.

      This antibody has been raised using a synthetic peptide derived from residues 250 to the C-terminus of Mouse Nanog. Your question seems to be about its specificity (species-specificity) and not its sensitivity.

      Since this antibody is not supposed to react with human Nanog, I would need to take your enquiry as a complaint. Would you be so kind to share some technical related details?

      I look forward to hearing from you soon.

      Read More

      Question

      Dear,

      I am a happy user of Abcam and today I have a question concerning Anti-Nanog antibody(ab14959).

      We start to optimize it using human tissue and we got a nice expression pattern on it. I just saw in your data sheet that this antibody does not react human tissue.

      However, I do believe its positivity in human tissue because it is so specific and reliable. Could you possible give your comments about this?

      Looking forward to hearining from you soon.

      Best Wishes,

      Read More
      Answer

      I am sorry to hear that you have been experiencing problems using this product in the application/species that you wish.

      Could you provide some further details of the protocol used and complete the following form (attached as a word document). It would be much appreciated if you could attach an image to the response.

      Thank you for your understanding and co-operation in this matter. I look forward to hearing from you soon and resolving this issue as soon as possible.

      Read More

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Nanog antibody

       Good
      Abreviews
      abreview image
      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
      Read More

      Mr. Carl Hobbs

      Verified customer

      Submitted Apr 12 2012

      Western blot abreview for Anti-Nanog antibody

       Good
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (ES cell)
      Loading amount
      10 µg
      Specification
      ES cell
      Treatment
      2000 U/ml mouse LIF constantly
      Gel Running Conditions
      Reduced Denaturing
      Blocking step
      Milk as blocking agent for 30 minute(s) · Concentration: 3% · Temperature: 20°C
      Read More

      Abcam user community

      Verified customer

      Submitted Feb 12 2009

      Question

      Customer bought Nanog antibody and would like an isotype control. Staining on ES cells.

      Read More
      Answer

      Thank you for your enquiry. There are at least three isotype controls you could use: normal rabbit serum (ab7487), ab27472, ab27478. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

      Read More

      Question

      What is the homology of your abs to bovine nanog (DQ069776)?

      Read More
      Answer

      Thank you for your enquiry and your patience in waiting for my reply. I have compared the immunogen sequences with the sequence of bovine nanog using a ClustalW alignment. For ab14959 less than 30% of the sequence is homologous, for ab21624 the homology is 50%, and for ab21603 the epitope is not known but the overall sequence homology is approximately 50%. The end result in my opinion is that the homology is not high enough for any of these antibodies to cross-react with bovine nanog. You would want to see a higher degree of homology to have any confidence that these antibodies would cross-react with bovine nanog. I hope this information helps, please do not hesitate to contact us if you need any more advice or information,

      Read More

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