Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: WB, IHC-P, ICC/IF, Flow Cyt, IP, ChIP
  • Reacts with: Mouse

Overview

  • Product name

    Anti-Nanog antibody [EPR20694] - BSA and Azide free
    See all Nanog primary antibodies

  • Description

    Rabbit monoclonal [EPR20694] to Nanog - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • IHC-P: Mouse E14.5 testis tissue.

  • General notes

    Ab231300 is the carrier-free version of ab214549. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab231300 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)
    Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214549).

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

    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 (blue), 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 (yellow). The immunoprecipitated DNA was quantified by real time PCR (SYBR green chemistry).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214549).

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

    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 1/5000 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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214549).

  • Flow Cytometry - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)
    Flow Cytometry - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)

    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.

     

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214549).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)

    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.

     This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214549).

    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] - BSA and Azide free (ab231300)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR20694] - BSA and Azide free (ab231300)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214549).

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

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

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