Overview

  • Product name

    Anti-MTH1 antibody [EPR15934]
    See all MTH1 primary antibodies
  • Description

    Rabbit monoclonal [EPR15934] to MTH1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Human MTH1 aa 50-150. The exact sequence is proprietary.
    Database link: P36639

  • Positive control

    • WB: Jurkat and A549 cell lysates. Human fetal thymus lysates. IHC: Human Spleen tissue. ICC/IF: PC-3 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: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR15934
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab197028 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 1/2000. Detects a band of approximately 18 kDa (predicted molecular weight: 23 kDa).
IHC-P 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.
ICC/IF 1/250.

Target

  • Function

    Antimutagenic. Acts as a sanitizing enzyme for oxidized nucleotide pools, thus suppressing cell dysfunction and death induced by oxidative stress. Hydrolyzes 8-oxo-dGTP, 8-oxo-dATP and 2-OH-dATP, thus preventing misincorporation of oxidized purine nucleoside triphosphates into DNA and subsequently preventing A:T to C:G and G:C to T:A transversions. Able to hydrolyze also the corresponding ribonucleotides, 2-OH-ATP, 8-oxo-GTP and 8-oxo-ATP. Does not play a role in U8 snoRNA decapping activity. Binds U8 snoRNA.
  • Tissue specificity

    Widely expressed with highest expression in thymus, testis, embryo and proliferating blood lymphocytes.
  • Sequence similarities

    Belongs to the Nudix hydrolase family.
    Contains 1 nudix hydrolase domain.
  • Developmental stage

    In peripheral blood lymphocytes, expressed at much higher levels in proliferating cells than in resting cells.
  • Post-translational
    modifications

    The N-terminus is blocked.
  • Cellular localization

    Cytoplasm. Mitochondrion matrix and Cytoplasm. Mitochondrion matrix. Nucleus. Mostly present in cytoplasm. Variant Met-124 has decreased efficiency in translocation to mitochondria.
  • Information by UniProt
  • Database links

  • Alternative names

    • 2-hydroxy-dATP diphosphatase antibody
    • 7 8 dihydro 8 oxoguanine triphosphatase antibody
    • 7 antibody
    • 8 oxo 7 8 dihydrodeoxyguanosine triphosphatase antibody
    • 8 oxo 7 8 dihydroguanosine triphosphatase antibody
    • 8 oxo dGTPase antibody
    • 8-dihydro-8-oxoguanine triphosphatase antibody
    • 8-oxo-dGTPase antibody
    • 8ODP_HUMAN antibody
    • MTH 1 antibody
    • MTH1 antibody
    • MutT human homolog 1 antibody
    • Nucleoside diphosphate linked moiety X motif 1 antibody
    • Nucleoside diphosphate linked moiety X type motif 1 antibody
    • Nucleoside diphosphate-linked moiety X motif 1 antibody
    • Nudix (nucleoside diphosphate linked moiety X) type motif 1 antibody
    • Nudix hydrolase 1 antibody
    • Nudix motif 1 antibody
    • Nudix type motif 1 antibody
    • NUDT 1 antibody
    • Nudt1 antibody
    see all

Images

  • Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: MTH1 knockout HAP1 cell lysate (20 µg)
    Lane 3: Jurkat cell lysate(20 µg)
    Lane 4: A549 cell lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab197028 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab197028 was shown to specifically react with MTH1 when MTH1 knockout samples were used. Wild-type and MTH1 knockout samples were subjected to SDS-PAGE. ab197028 at a dilution of 1/2000 and ab8245 (loading control to GAPDH) diluted at 1/10000 were incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded Human spleen tissue labeling MTH1 with ab197028 at 1/100 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasm and nuclear staining on Human spleen is observed. Counter stained with Hematoxylin. Negative control: Used PBS instead of primary antibody.

     

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

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized PC-3 cells (Human prostate cancer cell line)  labeling MTH1 with ab197028 at 1/250 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm and nuclear staining on PC-3 cell line is observed. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:-
    -ve control 1 - ab197028 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

  • Flow Cytometry analysis of PC-3 (human prostate adenocarcinoma) cells labeling MTH1 with purified ab188474 at 1/230 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)(1/2000 dilution) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • All lanes : Anti-MTH1 antibody [EPR15934] (ab197028) at 1/2000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate
    Lane 2 : A549 (Human lung carcinoma) whole cell lysate
    Lane 3 : Human fetal thymus lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Developed using the ECL technique.

    Predicted band size: 23 kDa
    Observed band size: 18 kDa
    why is the actual band size different from the predicted?


    Exposure time: 3 minutes


    The expression profile observed is consistent with what has been described in the literature (PMID: 11296483).The observed band may be isoform p18.

     

    Blocking/dilution buffer: 5% NFDM/TBST.

References

This product has been referenced in:

See 1 Publication for this product

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