Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-YTHDF2 antibody [EPR20318] (ab220163)

Overview

  • Product name

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

    Rabbit monoclonal [EPR20318] to YTHDF2
  • 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

    Synthetic peptide within Human YTHDF2 aa 250 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9Y5A9

  • 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

    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

    EPR20318
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab220163 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: 62 kDa.
IP 1/30.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IHC-P.
  • Target

    • Sequence similarities

      Contains 1 YTH domain.
    • Information by UniProt
    • Database links

    • Alternative names

      • 9430020E02Rik antibody
      • CAHL antibody
      • CLL associated antigen KW 14 antibody
      • CLL-associated antigen KW-14 antibody
      • HGRG8 antibody
      • High glucose regulated protein 8 antibody
      • High-glucose-regulated protein 8 antibody
      • NY REN 2 antibody
      • Renal carcinoma antigen NY REN 2 antibody
      • Renal carcinoma antigen NY-REN-2 antibody
      • YTH domain family 2 antibody
      • YTH domain family member 2 antibody
      • YTH domain family protein 2 antibody
      • YTH N6 methyladenosine RNA binding protein 2 antibody
      • YTHD2_HUMAN antibody
      • YTHDF 2 antibody
      • YTHDF2 antibody
      see all

    Images

    • 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.

    • 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.

    • 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.

    • 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.

       

    • 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.

    References

    ab220163 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (Spleen)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    20 µg
    Specification
    Spleen
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Linda Zhang

    Verified customer

    Submitted Jul 22 2019

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