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Developmental Biology Embryogenesis Embryonic stem cells Intracellular

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

Price and availability

284 121 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-Nanog antibody [EPR20694] - ChIP Grade (ab214549)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

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: ChIP, Flow Cyt, IHC-P, WB, ICC/IF, IP
  • Reacts with: Mouse

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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 & Species

    Application Species
    ChIP
    Mouse
    Flow Cyt
    Mouse
    ICC/IF
    Mouse
    IHC-P
    Mouse
    IP
    Mouse
    WB
    Mouse
    See all applications and species data
  • 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: 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

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

    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)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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