Overview

  • Product name

    Anti-YTHDC1 antibody [EPR21821]
    See all YTHDC1 primary antibodies
  • Description

    Rabbit monoclonal [EPR21821] to YTHDC1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
    Predicted to work with: Sheep, Goat, Horse, Guinea pig, Cow, Cat, Dog, Pig
  • Immunogen

    Recombinant fragment within Human YTHDC1 aa 300-550. The exact sequence is proprietary.
    Database link: Q96MU7

  • Positive control

    • WB: HeLa transfected with scrambled siRNA control whole cell lysate; Human brain tissue lysate; K562, C6, RAW 246.7, PC-12 and NIH/3T3 whole cell lysate. IHC-P: Human testis, endometrial carcinoma, mouse testis and rat spleen tissue.
  • 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

Applications

Our Abpromise guarantee covers the use of ab220159 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. Detects a band of approximately 95, 100 kDa (predicted molecular weight: 85 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    Images

    • Immunohistochemical analysis of paraffin-embedded mouse testis tissue stained for YTHDC1 with ab220159 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse testis (PMID: 27602518; PMID: 29103884) is observed. Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

      The section was incubated with ab220159 for 30 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded rat spleen tissue stained for YTHDC1 with ab220159 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat spleen (PMID: 27602518; PMID: 29103884) is observed. Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

      The section was incubated with ab220159 for 30 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded human endometrial carcinoma tissue stained for YTHDC1 with ab220159 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human endometrial carcinoma (PMID: 27602518; PMID: 29103884) is observed. Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

      The section was incubated with ab220159 for 30 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemical analysis of paraffin-embedded human testis tissue stained for YTHDC1 with ab220159 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human testis (PMID: 27602518; PMID: 29103884) is observed. Counterstained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). 

      The section was incubated with ab220159 for 30 mins at 37℃. The immunostaining staining was performed on a Leica Biosystems BOND® RX instrument.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • All lanes : Anti-YTHDC1 antibody [EPR21821] (ab220159) at 1/5000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate
      Lane 2 : YTHDC1 knockdown HeLa 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: 85 kDa
      Observed band size: 100 kDa
      why is the actual band size different from the predicted?


      Exposure time: 59 seconds


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

      The control and YTHDC1 knockdown cell lysates were kindly provided by an anonymous collaborator.

    • Lane 1 : Anti-YTHDC1 antibody [EPR21821] (ab220159) at 1/5000 dilution
      Lanes 2-5 : Anti-YTHDC1 antibody [EPR21821] (ab220159) at 1/1000 dilution

      Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 2 : C6 (rat glial tumor glial cell) whole cell lysate
      Lane 3 : RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 4 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate
      Lane 5 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 85 kDa
      Observed band size: 100,95 kDa why is the actual band size different from the predicted?



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

      The doublet observed is consistent with what has been shown in the literature (PMID: 26876937). The second band may represent the isoform 2 of YTHDC1 (PMID: 23765422).

      Exposure times:

      Lane 1:70 seconds;

      Lanes 2-5: 48 seconds.

    • Anti-YTHDC1 antibody [EPR21821] (ab220159) at 1/1000 dilution + Human brain tissue lysate at 20 µg

      Secondary
      VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution

      Predicted band size: 85 kDa
      Observed band size: 100,95 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


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

      The doublet observed is consistent with what has been shown in the literature (PMID: 26876937). The second band may represent the isoform 2 of YTHDC1 (PMID: 23765422). 

    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

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