Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-YTHDF1 antibody [EPR22349-41] (ab220162)

Submit a review Submit a question References (4)
Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
  • Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
  • Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
  • Immunoprecipitation - Anti-YTHDF1 antibody [EPR22349-4] (ab220162)
  • Anti-YTHDF1 antibody [EPR22349-41] (ab220162)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22349-41] to YTHDF1
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-YTHDF1 antibody [EPR22349-41]
    See all YTHDF1 primary antibodies

  • Description

    Rabbit monoclonal [EPR22349-41] to YTHDF1

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, IPmore details
    Unsuitable for: Flow Cyt,ICC/IF or IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human YTHDF1 aa 200 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9BYJ9

  • Positive control

    • WB: GST-tagged human YTHDF1 recombinant protein. Wild-type mESC whole cell lysate. HepG2, NIH/3T3 and HeLa whole cell lysate. Human brain lysate. Rat testis lysate. IP: YTHDF1 IP in HepG2 whole cell lysate.

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

    EPR22349-41

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab220162 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 1/1000. Predicted molecular weight: 61 kDa.
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.

    • Target

    Images

    • Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      All lanes : Anti-YTHDF1 antibody [EPR22349-41] (ab220162) at 1/1000 dilution

      Lane 1 : Wild-type mESC (mouse embryo stem cell) whole cell lysate
      Lane 2 : YTHDF1 knockout mESC whole cell lysate
      Lane 3 : HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate
      Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) 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

      Predicted band size: 61 kDa
      Observed band size: 61 kDa



      The wild-type and YTHDF1 knockout cell lysates were kindly provided by an anonymous collaborator.

      ab220162 was shown to specifically react with YTHDF1 in wild-type mESC cells as signal was lost in YTHDF1 knockout cells. Wild-type and YTHDF1 knockout samples were subjected to SDS-PAGE. ab220162 and ab181602 (Rabbit anti-GAPDH loading control) were incubated 1 hour at room temperature at 1/1000 dilution and 1/200,000 dilution respectively. Blots were developed with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) secondary antibody at 1/100,000 dilution for 1 hour at room temperature before imaging. The blot was developed on a BIO-RAD® ChemiDoc™ MP instrument using the ECL technique.

      Exposure times: Lanes 1 and 2: 59 seconds; Lanes 3 and 4: 26 seconds.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      All lanes : Anti-YTHDF1 antibody [EPR22349-41] (ab220162) at 1/1000 dilution

      Lane 1 : GST-tagged human YTHDF1 recombinant protein, 20 ng
      Lane 2 : GST-tagged human YTHDF2 recombinant protein, 20 ng
      Lane 3 : GST-tagged human YTHDF3 recombinant protein, 20 ng

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

      Predicted band size: 61 kDa
      Observed band size: 87 kDa
      why is the actual band size different from the predicted?


      Exposure time: 3 seconds


      The YTHDF recombinant proteins were kindly provided by an anonymous collaborator.

      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      Western blot - Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      All lanes : Anti-YTHDF1 antibody [EPR22349-41] (ab220162) at 1/1000 dilution

      Lane 1 : Human brain lysate at 20 µg
      Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 10 µg
      Lane 3 : Rat testis lysate at 20 µg

      Secondary
      Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
      Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 61 kDa



      Recommend using fresh lysates to avoid target protein being degraded.

      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lanes 1 and 2: 3 minutes; Lane 3: 1 minute.

    • Immunoprecipitation - Anti-YTHDF1 antibody [EPR22349-4] (ab220162)
      Immunoprecipitation - Anti-YTHDF1 antibody [EPR22349-4] (ab220162)

      YTHDF1 was immunoprecipitated from 0.35 mg HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate with ab220162 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220162 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
      Lane 1: HepG2 whole cell lysate 10 µg (Input).
      Lane 2: ab220162 IP in HepG2 whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220162 in HepG2 whole cell lysate.
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 3 seconds.

      Lysate were made freshly and used in IP test immediately to minimize protein degradation. Incubation time was 2h.

    • Anti-YTHDF1 antibody [EPR22349-41] (ab220162)
      Anti-YTHDF1 antibody [EPR22349-41] (ab220162)

    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

    References (4)

    Publishing research using ab220162? Please let us know so that we can cite the reference in this datasheet.

    ab220162 has been referenced in 4 publications.

    • Ding H  et al. GNAS promotes inflammation-related hepatocellular carcinoma progression by promoting STAT3 activation. Cell Mol Biol Lett 25:8 (2020). WB ; Human . PubMed: 32123532
    • Jin D  et al. m6A demethylase ALKBH5 inhibits tumor growth and metastasis by reducing YTHDFs-mediated YAP expression and inhibiting miR-107/LATS2-mediated YAP activity in NSCLC. Mol Cancer 19:40 (2020). WB ; Human . PubMed: 32106857
    • Zhao W  et al. METTL3 Facilitates Oral Squamous Cell Carcinoma Tumorigenesis by Enhancing c-Myc Stability via YTHDF1-Mediated m6A Modification. Mol Ther Nucleic Acids 20:1-12 (2020). WB ; Human . PubMed: 32145676
    • Huang B  et al. Cyclophosphamide Regulates N6-Methyladenosine and m6A RNA Enzyme Levels in Human Granulosa Cells and in Ovaries of a Premature Ovarian Aging Mouse Model. Front Endocrinol (Lausanne) 10:415 (2019). PubMed: 31316467

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    There are currently no Customer reviews or Questions for ab220162.
    Please use the links above to contact us or submit feedback about this product.

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