Validated using a knockout cell line
Recombinant
RabMAb

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

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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

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

    This product is a recombinant rabbit monoclonal antibody.

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

    Preservative: 0.01% Sodium azide
    Constituents: PBS, 40% Glycerol, 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.

    References

    This product has been referenced in:

    • 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). Read more (PubMed: 31316467) »
    See 1 Publication for this product

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

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