Astana Biomed Group, an authorized Abcam distributor in Central Asia

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, Recombinant fragment

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 & Species

    Application Species
    IP
    Human
    WB
    Mouse
    Rat
    Human
    Recombinant fragment
    See all applications and species data

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • 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.

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

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)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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