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Validated using a knockout cell lineRecombinantRabMAb

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

Reviews (4) Submit a question References (10)
Western blot - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Western blot - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunoprecipitation - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Flow Cytometry (Intracellular) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
  • Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR21820-49] to YTHDC2
  • Suitable for: Flow Cyt (Intra), IHC-P, IP, WB, IHC-Fr
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

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: Flow Cyt (Intra), IHC-P, IP, WB, IHC-Frmore details
    Unsuitable for: ICC/IF

  • Species reactivity

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

  • Immunogen

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

  • 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. Flow Cyt (intra): HeLa cells. IP: HeLa whole cell lysate. IHC-Fr: Mouse testis, Rat testis tissues.

  • 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

    EPR21820-49

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/600.
IHC-P (1)
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
WB (2)
1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).
IHC-Fr
1/50.
Notes
Flow Cyt (Intra)
1/600.
IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
WB
1/1000. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).
IHC-Fr
1/50.
Application notes
Is unsuitable for ICC/IF.

Target

Images

  • Western blot - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Western blot - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    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.

  • Western blot - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Western blot - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    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.

  • Immunoprecipitation - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunoprecipitation - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    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).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    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 Cytometry (Intracellular) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Flow Cytometry (Intracellular) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    Intracellular 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. 

     

     

  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised mouse testis tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). Showing cytoplasmic staining on mouse testis. Nuclear counterstain: DAPI. 

  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    Negative control: (PMID: 29087293)

    Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised mouse heart tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). No staining observed on mouse heart. Nuclear counterstain: DAPI. 

  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised rat testis tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). Showing cytoplasmic staining on rat testis. Nuclear counterstain: DAPI. 

  • Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)
    Immunohistochemistry (Frozen sections) - Anti-YTHDC2 antibody [EPR21820-49] (ab220160)

    Negative control: (PMID: 29087293)

    Immunohistochemical analysis of 4% paraformaldehyde fixed, 0.2% Triton X-100 permeabilised rat heart tissue labeling YTHDC2 with ab220160 at 1/50 dilution (11 ug/mL). Followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary antibody at 1/1000 dilution (2 ug/mL). No staining observed on rat heart. Nuclear counterstain: DAPI. 

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

Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

References (10)

Publishing research using ab220160? Please let us know so that we can cite the reference in this datasheet.

ab220160 has been referenced in 10 publications.

  • Jiao Y  et al. The m6A reader YTHDC2 promotes SIRT3 expression by reducing the stabilization of KDM5B to improve mitochondrial metabolic reprogramming in diabetic peripheral neuropathy. Acta Diabetol 60:387-399 (2023). WB, RIP ; Rat . PubMed: 36574062
  • Saito Y  et al. YTHDC2 control of gametogenesis requires helicase activity but not m6A binding. Genes Dev N/A:N/A (2022). WB, CLIP ; Mouse . PubMed: 35058317
  • Ding Y  et al. Circular RNA midline-1 (circMID1) promotes proliferation, migration, invasion and glycolysis in prostate cancer. Bioengineered 13:6293-6308 (2022). WB, RIP ; Human . PubMed: 35212614
  • Qian B  et al. RNA binding protein RBM46 regulates mitotic-to-meiotic transition in spermatogenesis. Sci Adv 8:eabq2945 (2022). WB, RIP ; Mouse . PubMed: 36001654
  • Tao L  et al. FTO modifies the m6A level of MALAT and promotes bladder cancer progression. Clin Transl Med 11:e310 (2021). PubMed: 33634966
  • Shen M  et al. N6-methyladenosine modification regulates ferroptosis through autophagy signaling pathway in hepatic stellate cells. Redox Biol 47:102151 (2021). WB ; Rat, Human . PubMed: 34607160
  • Zhou X  et al. Analysis of Genetic Alteration Signatures and Prognostic Values of m6A Regulatory Genes in Head and Neck Squamous Cell Carcinoma. Front Oncol 10:718 (2020). PubMed: 32547941
  • Li Y  et al. The m6A reader protein YTHDC2 is a potential biomarker and associated with immune infiltration in head and neck squamous cell carcinoma. PeerJ 8:e10385 (2020). IHC-P ; Human . PubMed: 33304653
  • Sun S  et al. Downregulation of m6 A reader YTHDC2 promotes tumor progression and predicts poor prognosis in non-small cell lung cancer. Thorac Cancer 11:3269-3279 (2020). PubMed: 32956555
  • Toyo-oka T  et al. Temporary salvage of ischemic myocardium by the protease inhibitor bis[ethyl(2R,3R)-3-[(S)-methyl-1-[4-(2,3,4-trimethoxyphenyl-methyl) piperazin-1-ylcarbonyl]butyl-carbonyl]oxiran-2-carbo xylate]sulfate. Arzneimittelforschung 36:671-5 (1986). WB ; Human . PubMed: 3521614

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-YTHDC2 antibody [EPR21820-49]

 Excellent
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Aug 30 2019

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