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RecombinantRabMAb

Recombinant Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

  • Datasheet
  • SDS
  • Certificate of Compliance
Reviews (3) Submit a question References (4)

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ChIP - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Flow Cytometry (Intracellular) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Immunoprecipitation - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20694] to Nanog - ChIP Grade
  • Suitable for: Flow Cyt (Intra), ChIP, IHC-P, WB, ICC/IF, IP
  • Reacts with: Mouse

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free

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Recombinant human Nanog protein (ab134886)
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Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)

View more associated products

Overview

  • Product name

    Anti-Nanog antibody [EPR20694] - ChIP Grade
    See all Nanog primary antibodies
  • Description

    Rabbit monoclonal [EPR20694] to Nanog - ChIP Grade
  • Host species

    Rabbit
  • Tested applications

    Suitable for: Flow Cyt (Intra), ChIP, IHC-P, WB, ICC/IF, IPmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: F9 and ES-D3 whole cell lysates. IHC-P: Mouse E14.5 testis tissue. ICC/IF: F9 cells. Flow Cyt (intra): F9 cells. IP: F9 whole cell lysate. ChIP: Chromatin prepared from F9 cells.
  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR20694
  • Isotype

    IgG
  • Research areas

    • Stem Cells
    • Embryonic Stem Cells
    • Intracellular
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • SMADs
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cancer
    • Cell cycle
    • Cell differentiation
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies
    • Developmental Biology
    • Embryogenesis
    • Embryonic stem cells
    • Surface molecules

Associated products

  • Alternative Versions

    • Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Recombinant Protein

    • Recombinant human Nanog protein (ab134886)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab214549 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 (Intra)
1/60.
ChIP
Use 5 µg for 25 µg of chromatin.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 29-42 kDa (predicted molecular weight: 34 kDa).
ICC/IF (1)
1/100.
IP
1/30.
Notes
Flow Cyt (Intra)
1/60.
ChIP
Use 5 µg for 25 µg of chromatin.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000. Detects a band of approximately 29-42 kDa (predicted molecular weight: 34 kDa).
ICC/IF
1/100.
IP
1/30.

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.
  • Target information above from: UniProt accession Q9H9S0 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 71950 Mouse
    • SwissProt: Q80Z64 Mouse
    • Unigene: 6047 Mouse
    • 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

    • ChIP - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      ChIP - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

      Chromatin was prepared from F9 cells according to the Abcam X-ChIP protocol. Cells were fixed with 1% formaldehyde for 10 minutes. The ChIP was performed with 25 µg of chromatin, 5µg of ab214549 (red), and 20 µl of protein A/G sepharose beads slurry (10 µl of sepharose A beads + 10 µl of sepharose G beads). Then 5 μg of rabbit normal IgG was added to the control beads (grey). The immunoprecipitated DNA was quantified by real time PCR (SYBR green chemistry).

    • Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      All lanes : Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549) at 1/1000 dilution

      Lane 1 : F9 (Mouse embryonic testicular cancer cell line) whole cell lysate
      Lane 2 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 34 kDa
      Observed band size: 29-42 kDa why is the actual band size different from the predicted?


      Exposure time: 70 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The multiple bands observed are consistent with the literature (PMID: 24936455). Negative control: NIH/3T3 (PMID: 12787505).

    • Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Western blot - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549) at 1/1000 dilution + ES-D3 (mouse embryonic multipotent stem cell line) whole cell lysate at 10 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Developed using the ECL technique.

      Predicted band size: 34 kDa
      Observed band size: 29-42 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The multiple bands observed are consistent with the literature (PMID: 24936455).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

      Immunohistochemical analysis of paraffin-embedded mouse E14.5 testis tissue labeling Nanog with ab214549 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Mainly nuclear staining is observed in testis of mouse embryo E14.5 (PMID: 15939376; PMID: 12787505). Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

      Immunohistochemical analysis of paraffin-embedded adult mouse testis tissue labeling Nanog with ab214549 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Negative tissue: No staining on adult mouse testis is observed(PMID: 12787505; PMID: 15939376).

      Counter stained with Hematoxylin.

      Heat mediated antigen retrieval was performed with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeablized F9 (mouse embryonic testicular cancer cell line) and NIH/3T3 (mouse embryo fibroblast cell line) cells labeling Nanog with ab214549 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in F9 cell line. Negative control: NIH/3T3 (PMID: 17352742).

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Flow Cytometry (Intracellular) - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

      Intracellular flow cytometric analysis of 4 % paraformaldehyde-fixed, 90% methanol permeabilized F9 (mouse embryonic testicular cancer cell line, Right) and NIH/3T3 (mouse embryo fibroblast cell line, Left) cells labeling Nanog with ab214549 at 1/60 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. 

      Negative control: NIH/3T3 cell line. 

       

       

       

    • Immunoprecipitation - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Immunoprecipitation - Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

      Nanog was immunoprecipitated from 0.35 mg of F9 (mouse embryonic testicular cancer cell line) whole cell lysate with ab214549 at 1/1000 dilution. Western blot was performed from the immunoprecipitate using ab214549 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution

      Lane 1: F9 whole cell lysate 10 μg (input)

      Lane 2: ab214549 IP in F9 whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214549 in F9 whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 15 seconds.

      The multiple bands observed are consistent with the literature (PMID: 24936455).

    • Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
      Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    Certificate of Compliance

    To download a Certificate of Compliance, please enter your Lot number below:

    References (4)

    Publishing research using ab214549? Please let us know so that we can cite the reference in this datasheet.

    ab214549 has been referenced in 4 publications.

    • Tan A  et al. TFEB regulates pluripotency transcriptional network in mouse embryonic stem cells independent of autophagy-lysosomal biogenesis. Cell Death Dis 12:343 (2021). PubMed: 33795648
    • Li J  et al. Aberrant Gcm1 expression mediates Wnt/ß-catenin pathway activation in folate deficiency involved in neural tube defects. Cell Death Dis 12:234 (2021). PubMed: 33664222
    • Zhu Q  et al. O-GlcNAcylation regulates the methionine cycle to promote pluripotency of stem cells. Proc Natl Acad Sci U S A 117:7755-7763 (2020). PubMed: 32193337
    • Zhang S  et al. The Hippo Effector Transcriptional Coactivator with PDZ-Binding Motif Cooperates with Oncogenic ß-Catenin to Induce Hepatoblastoma Development in Mice and Humans. Am J Pathol 190:1397-1413 (2020). PubMed: 32283103

    Customer reviews and Q&As

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    1-3 of 3 Abreviews or Q&A

    IHC - Wholemount abreview for Anti-Nanog antibody [EPR20694] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (120h Blimp1-GFP gastruloid)
    Specification
    120h Blimp1-GFP gastruloid
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Abcam user community

    Verified customer

    Submitted Mar 09 2023

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Nanog antibody [EPR20694] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cultured Cells (mouse embryonic stem cells)
    Specification
    mouse embryonic stem cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°C
    Fixative
    Methanol
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Malgorzata Borkowska

    Verified customer

    Submitted Apr 14 2019

    IHC - Wholemount abreview for Anti-Nanog antibody [EPR20694] - ChIP Grade

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    IHC - Wholemount
    Sample
    Mouse Tissue (whole mount E9.5 mouse embryo)
    Specification
    whole mount E9.5 mouse embryo
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Malgorzata Borkowska

    Verified customer

    Submitted Apr 04 2019

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