Product nameAnti-MTH1 antibody [EPR15934]
See all MTH1 primary antibodies
DescriptionRabbit monoclonal [EPR15934] to MTH1
Tested applicationsSuitable for: WB, IHC-P, Flow Cyt, ICC/IFmore details
Species reactivityReacts with: Human
Predicted to work with: Mouse, Rat
Recombinant fragment within Human MTH1 aa 50-150. The exact sequence is proprietary.
Database link: P36639
- WB: Jurkat and A549 cell lysates. Human fetal thymus lysates. IHC: Human Spleen tissue. ICC/IF: PC-3 cells.
Storage instructionsShipped 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 bufferPreservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
PurityProtein A purified
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.
|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.|
FunctionAntimutagenic. 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 specificityWidely expressed with highest expression in thymus, testis, embryo and proliferating blood lymphocytes.
Sequence similaritiesBelongs to the Nudix hydrolase family.
Contains 1 nudix hydrolase domain.
Developmental stageIn peripheral blood lymphocytes, expressed at much higher levels in proliferating cells than in resting cells.
modificationsThe N-terminus is blocked.
Cellular localizationCytoplasm. Mitochondrion matrix and Cytoplasm. Mitochondrion matrix. Nucleus. Mostly present in cytoplasm. Variant Met-124 has decreased efficiency in translocation to mitochondria.
- Information by UniProt
- 2-hydroxy-dATP diphosphatase antibody
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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.
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.