Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-YTHDF2 antibody [EPR20318] (ab220163)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20318] to YTHDF2
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, Recombinant fragment

Overview

  • Product name

    Anti-YTHDF2 antibody [EPR20318]
    See all YTHDF2 primary antibodies

  • Description

    Rabbit monoclonal [EPR20318] to YTHDF2

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: Wild-type mESC, HEK-293T, HT-1080 treated with 10µM MG-132 for 4 hours, K562, HEK-293 and HeLa whole cell lysate. Human brain, mouse and rat testis tissue lysate; GST-tagged human YTHDF2 recombinant protein. IP: HepG2 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

    EPR20318

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/1000 dilution

    Lane 1 : Wild-type mESC (mouse embryo stem cell) whole cell lysate
    Lane 2 : YTHDF2 knockout mESC whole cell lysate
    Lane 3 : 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: 62 kDa


    Exposure time: 48 seconds


    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

    The wild-type and YTHDF2 knockout cell lysates were kindly provided by an anonymous collaborator.

    ab220163 was shown to specifically react with YTHDF2 in wild-type mESC cells as signal was lost in YTHDF2 knockout cells. Wild-type and YTHDF2 knockout samples were subjected to SDS-PAGE. ab220163 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.

  • Immunoprecipitation - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    Immunoprecipitation - Anti-YTHDF2 antibody [EPR20318] (ab220163)

    ab220163 Immunoprecipitating YTHDF2 in human HepG2 whole cell lysate. 10 µg of cell lysate was incubated with primary antibody (1/30). For western blotting ab220163 (1/1000) was used to confirm successful immunoprecipitation. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at (1/5000). For lane 3 Rabbit monoclonal IgG (ab172730) was used instead of ab220163.

    In panel A, the lysate was stored at -80oC prior to the IP and incubation with the primary antibody was carried out overnight. In panel B, lysate was freshly made and used in the IP test immediately to minimize protein degradation. The incubation time was also shortened from overnight to 2h.

  • Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/5000 dilution

    Lane 1 : Untreated HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate at 20 µg
    Lane 2 : HT-1080 treated with 10µM MG-132 (ab141003) for 4 hours, whole cell lysate at 10 µg

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

    Predicted band size: 62 kDa


    Exposure time: 103 seconds


    Blocking/diluting buffer and concentration: 5% NFDM/TBST.

  • Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/1000 dilution

    Lane 1 : Human brain tissue lysate
    Lane 2 : Mouse testis tissue lysate
    Lane 3 : Rat testis tissue lysate
    Lane 4 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
    Lane 5 : K562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Lane 1 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
    Lanes 2-5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 62 kDa



    Blocking/diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: Lane 1: 3 minutes; Lane 2: 92 seconds; Lane 3: 37 seconds; Lane 4: 8 seconds; Lane 5: 15 seconds.

     

  • Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    Western blot - Anti-YTHDF2 antibody [EPR20318] (ab220163)
    All lanes : Anti-YTHDF2 antibody [EPR20318] (ab220163) at 1/1000 dilution

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

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

    Predicted band size: 62 kDa


    Exposure time: 3 seconds


    Blocking/diluting buffer and concentration: 5% NFDM/TBST.

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

  • Anti-YTHDF2 antibody [EPR20318] (ab220163)
    Anti-YTHDF2 antibody [EPR20318] (ab220163)

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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