Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23544-19] to YTHDF2
  • Suitable for: Flow Cyt, WB, IHC-P, IP, ICC
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-YTHDF2 antibody [EPR23544-19]
    See all YTHDF2 primary antibodies

  • Description

    Rabbit monoclonal [EPR23544-19] to YTHDF2

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Mouse
    Human
    ICC
    Mouse
    Human
    IHC-P
    Mouse
    Rat
    Human
    IP
    Mouse
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HeLa, LNCaP, MEF and PC-12 whole cell lysates; His-tagged human YTHDF2 recombinant protein. IHC-P: Human testis tissue; Mouse testis tissue; Rat testis tissue. ICC: HeLa and NIH/3T3 cells. Flow cyt: HeLa and NIH/3T3 cells. IP: HeLa and NIH/3T3 whole cell lysates.

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

    EPR23544-19

  • Isotype

    IgG

  • Research areas

Images

  • Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial adenocarcinoma cell line) (siRNA control) and HeLa (siRNA YTHDF) cells labelling YTHDF2 with ab246514 at 1/100 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in HeLa cell line transfected with scramble siRNA and reduced staining in HeLa cell line transfected with siRNA YTHDF2. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunocytochemistry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embryo fibroblast cell line) cells labelling YTHDF2 with ab246514 at 1/50 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

  • Flow Cytometry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Flow Cytometry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling YTHDF2 with ab246514 at 1/500 dilution (0.1ug) (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.

  • Immunoprecipitation - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunoprecipitation - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    YTHDF2 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast cell line) whole cell lysate with ab246514 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246514 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: NIH/3T3 whole cell lysate 10 ug

    Lane 2: ab246514 IP in NIH/3T3 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246514 in NIH/3T3 whole cell lysate

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

    Exposure time: 84 seconds.

    Lysates were made freshly and used in IP test immediately to minimize protein degradation.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling YTHDF2 with ab246514 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in mouse testis (PMID: 30903744, PMID: 29103884). The section was incubated with ab246514 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Western blot - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Western blot - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    All lanes : Anti-YTHDF2 antibody [EPR23544-19] (ab246514) at 1/1000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 2 : LNCaP (human prostate carcinoma epithelial cell) whole cell lysate
    Lane 3 : MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate
    Lane 4 : 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), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 62 kDa
    Observed band size: 62 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure times: Lanes 1-2: 37 seconds; Lane 3: 180 seconds; Lane 4:37 seconds.

    The molecular mass observed is consistent with the literature (PMID: 28104805).

    Lysates should be made freshly and used in WB immediately to minimize protein degradation.

  • Western blot - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Western blot - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    All lanes : Anti-YTHDF2 antibody [EPR23544-19] (ab246514) at 1/5000 dilution

    Lane 1 : LIF-His-tagged human YTHDF1 recombinant protein, 10 ng
    Lane 2 : His-tagged human YTHDF2 recombinant protein, 10 ng
    Lane 3 : His-tagged human YTHDF3 recombinant protein, 10 ng

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 62 kDa
    Observed band size: 62 kDa



    Blocking and dilution buffer: 5% NFDM/TBST.

    Exposure time: 59 seconds.

    These rec proteins were made in house.
    LIF-His-tagged human YTHDF1 recombinant protein was expressed from a mammalian - HEK-293 expression system.
    His-tagged human YTHDF2 recombinant protein is extracts of E.coli expressing YTHDF2.
    His-tagged human YTHDF3 recombinant protein is extracts of E.coli expressing YTHDF3.

     

  • Immunoprecipitation - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunoprecipitation - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    YTHDF2 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell line) whole cell lysate with ab246514 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab246514 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HeLa whole cell lysate 10 ug

    Lane 2: ab246514 IP in HeLa whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab246514 in HeLa whole cell lysate

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

    Exposure time: 10 seconds.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Immunohistochemical analysis of paraffin-embedded Rat testis tissue labeling YTHDF2 with ab246514 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in rat testis (PMID: 30903744, PMID: 29103884). The section was incubated with ab246514 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Flow Cytometry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Flow Cytometry - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast cell line) cells labelling YTHDF2 with ab246514 at 1/500 dilution (0.1ug) (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 (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

    Immunohistochemical analysis of paraffin-embedded Human testis tissue labeling YTHDF2 with ab246514 at 1/2000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic staining in human testis (PMID: 30903744, PMID: 29103884).The section was incubated with ab246514 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Anti-YTHDF2 antibody [EPR23544-19] (ab246514)
    Anti-YTHDF2 antibody [EPR23544-19] (ab246514)

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