Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

Overview

  • Product name

    Anti-YTHDC2 antibody [EPR21820-49]
    See all YTHDC2 primary antibodies
  • Description

    Rabbit monoclonal [EPR21820-49] to YTHDC2
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, IP, WBmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Cow, Pig, Non human primates
  • Immunogen

    Recombinant fragment within Human YTHDC2 aa 1100 to the C-terminus. The exact sequence is proprietary.
    Database link: Q9H6S0

  • Positive control

    • WB: Wild-type mESC, HT-1080, HeLa, HEK-293, NIH/3T3, PC-12 whole cell lysates. IHC-P: Human testis, Mouse testis and Rat testis tissues. FC: HeLa cells. IP: HeLa 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

    EPR21820-49
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab220160 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
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/600.
IP 1/30.
WB 1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).
  • Application notes
    Is unsuitable for ICC/IF.
  • Target

    Images

    • All lanes : Anti-YTHDC2 antibody [EPR21820-49] (ab220160) at 1/1000 dilution

      Lane 1 : Wild-type mESC (mouse embryo stem cell) whole cell lysate
      Lane 2 : YTHDC2 knockout mESC whole cell lysate

      Lysates/proteins at 40 µg per lane.

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

      Predicted band size: 160 kDa
      Observed band size: 160 kDa


      Exposure time: 3 minutes


      Blocking and dilution bufer: 5% NFDM/TBST.

      The wild-type and YTHDC2 knockout cell lysates were kindly provided by an anonymous collaborator.

      ab220160 was shown to specifically react with YTHDC2 in wild-type mESC cells as signal was lost in YTHDC2 knockout cells. Wild-type and YTHDC2 knockout samples were subjected to SDS-PAGE. ab220160 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.

    • All lanes : Anti-YTHDC2 antibody [EPR21820-49] (ab220160) at 1/1000 dilution

      Lane 1 : HT-1080 (human fibrosarcoma epithelial cell) whole cell lysate
      Lane 2 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 3 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate
      Lane 4 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
      Lane 5 : PC-12 (rat adrenal gland pheochromocytoma) 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: 160 kDa
      Observed band size: 160 kDa



      Blocking and dilution buffer: 5% NFDM/TBST.

      Exposure times.

      Lane 1: 3 minutes; Lanes 2-3: 37 seconds; Lane 4: 3 minutes; Lane 5: 37 seconds.

    • YTHDC2 was immunoprecipitated from 0.35 mg of HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab220160 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab220160 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

      Lane 1: HeLa whole cell lysate 10 μ (Input).
      Lane 2: ab220160IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab220160 in HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 30 seconds.

    • Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling YTHDC2 with ab220160 at 1/500 dilution (1.19 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on rat testis (PMID:28380054). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling YTHDC2 with ab220160 at 1/500 dilution (1.19 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on mouse testis (PMID:28380054). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling YTHDC2 with ab220160 at 1/500 dilution (1.19 ug/ml) followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on human testis (PMID:28380054). Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling YTHDC2 with ab220160 at 1/600 (Red) compared with a Rabbit monoclonal IgG (ab172730) / Black isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

    References

    ab220160 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    1-3 of 3 Abreviews or Q&A

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

    Linda Zhang

    Verified customer

    Submitted Oct 16 2019

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (testis)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    40 µg
    Specification
    testis
    Blocking step
    Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Aug 30 2019

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (testis)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate antigen repair solution
    Permeabilization
    Yes - PBST (0.5% Triton X-100 in PBS)
    Specification
    testis
    Blocking step
    BSA as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Fixative
    Formaldehyde

    Abcam user community

    Verified customer

    Submitted Aug 30 2019

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