MTH1 Antibody - BSA Free

Knockout Validated: MTH1 Antibody [NB100-109] - Western blot shows lysates of HEK293T human embryonic kidney parental cell line and MTH1 knockout (KO) HEK293T cell line. PVDF membrane was probed with 1:500 of Rabbit Anti-Human MTH1 Polyclonal Antibody (Catalog # NB100-109) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (Catalog #HAF008). Specific band was detected for MTH1 at approximately 18 kDa (as indicated) in the parental HEK293T cell line, but is not detectable in the knockout HEK293T cell line. This experiment was conducted under reducing conditions.

Simple Western: MTH1 Antibody [NB100-109] - Simple Western lane view shows a specific band for MTH1 in 0.5 mg/ml of A431 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Western Blot: MTH1 Antibody [NB100-109] - The cells from esophageal cancer cell lines: KYSE-450, EC109 and EC9706 (d, e), liver cancer cell lines: SMMC-7721, HepG2 and ZIP177 (f, g), gastric cancer cell lines: MGC-803, HGC-27, SGC-7901 and MKN45 (h, i), as well as the corresponding normal cell lines: Het-1A (d, e), L02 (f, g) and GES-1 (h, i) were cultured and lysed. The MTH1 protein levels were determined by Western Blot. GAPDH was used as a loading control. At least three independent experiments were performed for each group. j The cells indicated above were lysed and the total mRNA was extracted. The mRNA level of MTH1 was determined by RT-PCR. GAPDH was used as a control. At least three independent experiments were performed for each group. Image collected and cropped by Citeab from the following publication (Potent and specific MTH1 inhibitors targeting gastric cancer. Cell Death Dis (2019)) licensed under a CC-BY license.

Western Blot: MTH1 Antibody [NB100-109] - The antioxidant pathways are activated during EBV infection and are required for growth transformation. Freshly isolated B-lymphocytes infected with the transforming B95-8 strain of EBV were cultured for up to 2 weeks in the presence or absence of MTH1 inhibitors. Protein expression was monitored by western blots, cell proliferation and activation of the DDR were assessed by 3H-Thy incorporation and staining for I3H2AX, respectively. Representative western blots illustrating the expression of MTH1, MUTYH and OGG1 in ex vivo untreated B-cell and freshly EBV infected and SAC induced B blasts cultured for comparable times and showing similar levels of cell proliferation. Image collected and cropped by Citeab from the following publication (The Epstein-Barr virus nuclear antigen-1 upregulates the cellular antioxidant defense to enable B-cell growth transformation and immortalization. Oncogene (2020)) licensed under a CC-BY license.

Immunocytochemistry/Immunofluorescence: MTH1 Antibody [NB100-109] - HeLa cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X PBS + 0.05% Triton-X100. The cells were incubated with anti-MTH1 at 2 ug/ml overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Western Blot: MTH1 Antibody [NB100-109] - EBNA1 promotes the upregulation of oxidative DNA damage repair pathways in EBV converted cell lines and EBV positive BLs. Densitometry quantification of the specific bands. The intensity of the specific band in EBV positive cells relative the EBV-negative parental is shown. Image collected and cropped by Citeab from the following publication (The Epstein-Barr virus nuclear antigen-1 upregulates the cellular antioxidant defense to enable B-cell growth transformation and immortalization. Oncogene (2020)) licensed under a CC-BY license.

Western Blot: MTH1 Antibody [NB100-109] - MTH1 antibody was tested in the labeled lanes.

Western Blot: MTH1 Antibody [NB100-109] - Analysis of MTH1 in 1. Ntera2, 2. MCF7 cell lysate, 3. A431 cell lysate and 4. COS7 cell lysate.

Immunocytochemistry/Immunofluorescence: MTH1 Antibody [NB100-109] - A431 cells were fixed for 10 minutes using 10% formalin and then permeabilized for 5 minutes using 1X TBS + 0.5% Triton-X100. The cells were incubated with anti-MTH1 (NB100-109) at a 1:200 dilution overnight at 4C and detected with an anti-rabbit Dylight 488 (Green) at a 1:500 dilution. Alpha tubulin was used as a co-stain at a 1:1000 dilution and detected with an anti-mouse Dylight 550 (Red) at a 1:500 dilution. Nuclei were counterstained with DAPI (Blue). Cells were imaged using a 40X objective.

Knockdown Validated: MTH1 Antibody [NB100-109] - MTH1 suppression reduces the two gastric cancer cell survival. MGC-803, HGC-27 and GES-1 cells were treated by MTH1 specific siRNA 1 and #2 for 72h. Non-targeting siRNA(NT) treatment was used as control. Western Blot analysis of the protein levels of MTH1, MUTYH, OGG1, p21, ATMpS1981 and p53pS15, after the cells were treated with siRNA#2. Densitometry shows relative protein expression normalized by GAPDH. Image collected and cropped by Citeab from the following publication (Potent and specific MTH1 inhibitors targeting gastric cancer. Cell Death Dis (2019)) licensed under a CC-BY license.

Knockdown Validated: MTH1 Antibody [NB100-109] - Simultaneous suppression of MTH1 and MYH was efficiently achieved using a two-vector system. Steps of cell line establishment are illustrated here. Top 15% of cells expressing GFP and RFP670 were sorted to improve the knockdown efficiency. Expression levels of MYH and MTH1 were analyzed after 96h of treatment with shRNA set2. Data were double normalized with GAPDH and NT-shRNA control samples and expressed as percentage. Image collected and cropped by Citeab from the following publication (hMYH and hMTH1 cooperate for survival in mismatch repair defective T-cell acute lymphoblastic leukemia. Oncogenesis (2016)) licensed under a CC-BY license.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, ICC/IF, IHC, WB, KD, KO, Single-Cell Western
• Clonality: Polyclonal
• Host: Rabbit
• Conjugate: Unconjugated
• Format: BSA Free
• Concentration: 1.0 mg/ml

MTH1 Antibody - BSA Free Summary

• Immunogen: A C-terminal peptide derived from human MTH1, conjugated to KLH. [UniProt# P36639]
• Isotype: IgG
• Clonality: Polyclonal
• Host: Rabbit
• Gene: NUDT1
• Purity: Immunogen affinity purified

Applications/Dilutions

• Dilutions:
  • Immunoblotting reported in scientific literature (PMID 21076467)
  • Immunocytochemistry/Immunofluorescence 1:50-1:200
  • Immunohistochemistry 1:200
  • Immunohistochemistry-Frozen 1:200
  • Immunohistochemistry-Paraffin 1:200
  • Knockdown Validated
  • Knockout Validated
  • Simple Western 1:50
  • Single Cell Western 100 ug/ml
  • Western Blot 1:500
• Application Notes: In WB, a band can be seen at approx. 18kDa.
  
  In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
  The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Theoretical MW: 18 kDa.
  Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
• Control:

  HeLa Whole Cell Lysate

Reactivity Notes

Rat reactivity reported in scientific literature (PMID: 12706856).

Packaging, Storage & Formulations

• Storage: Aliquot and store at -20C or -80C. Avoid freeze-thaw cycles.
• Buffer: PBS
• Preservative: 0.02% Sodium Azide
• Concentration: 1.0 mg/ml
• Purity: Immunogen affinity purified

Alternate Names for MTH1 Antibody - BSA Free

• 8-oxo-7,8-dihydrodeoxyguanosine triphosphatase
• 8-oxo-dGTPase
• EC 3.6.1.-
• EC 3.6.1.56
• MTH1
• MTH1Nucleoside diphosphate-linked moiety X motif 1
• mutT human homolog 1,8-oxo-dGTPase
• nucleoside diphosphate-linked moiety X-type motif 17,8-dihydro-8-oxoguanine triphosphatase
• nudix (nucleoside diphosphate linked moiety X)-type motif 1,8-oxo-7,8-dihydroguanosine triphosphatase
• Nudix motif 1
• NUDT1

Background

MTH1 (human MutT Homolog 1) is an oxidized purine nucleoside triphosphatase enzyme that belongs to the Nudix hydrolase family and it acts as major mammalian detoxifier of the oxidized DNA precursor, 8-oxo-dGTP thereby exerting antimutagenic effects and suppressing cell dysfunction/death induced by oxidative stress. MTH1 hydrolyzes 8-oxo-dGTP, 8-oxo-dATP and 2-OH-dATP, thereby preventing misincorporation of oxidized purine nucleoside triphosphates into DNA and subsequently preventing A:T to C:G and G:C to T:A transversions. MTH1 also hydrolyzes the corresponding ribonucleotides, 2-OH-ATP, 8-oxo-GTP and 8-oxo-ATP. 2-OH-dADP, 8-OH-dGDP and 8-OH-dGTP inhibit the 2-OH-dATPase activity whereas 8-OH-dGTPase activity is inhibited by 8-OH-dGDP, 2-OH-dADP and 2-OH-dATP. Localized mainly in the cytoplasm, MTH1 has been found in nucleus and mitochondria also, and is widely expressed with highest expression in thymus, testis, embryo, proliferating blood lymphocytes etc. Oxidative DNA damage, e.g. 8-oxoG is known to accumulate both in nuclear and mitochondrial genomes during aging, carcinogenesis or various neurodegenerative diseases, such as Parkinson's disease, Alzheimer's disease or amyotrophic lateral sclerosis and MTH1 deficiency can increase susceptibility to these oxidative damage mediated dysfunctions.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for MTH1 Antibody (NB100-109)(32)

Publications using NB100-109

• Henriksson S, CalderOn-MontaNo J, Solvie D et al. Overexpressed C-Myc Sensitize Cells to TH1579, an Mitotic Arrest and Oxidative DNA Damage Inducer Preprint 2022-08-22 [PMID: 36551206] (WB, Human)
• Bialkowski K, Szpila A Specific 8-oxo-dGTPase activity of MTH1 (NUDT1) protein as a quantitative marker and prognostic factor in human colorectal cancer Free radical biology & medicine 2021-10-05 [PMID: 34624481]
• Karsten S, Fiskesund R, Zhang XM Et al. MTH1 as a target to alleviate T cell driven diseases by selective suppression of activated T cells Cell death and differentiation 2021-08-27 [PMID: 34453118] (WB, Human)
• Zhang L, Misiara L, Samaranayake GJ, et al. OGG1 co-inhibition antagonizes the tumor-inhibitory effects of targeting MTH1 Redox biology 2021-01-02 [PMID: 33450725] (WB, Human)
• Arczewska KD, Krasuska W, Stachurska A et al. hMTH1 and GPX1 expression in human thyroid tissue is interrelated to prevent oxidative DNA damage DNA Repair (Amst.) 2020-08-20 [PMID: 32877752] (WB, Human)
• Godoy PRDV, Pour Khavari A, Rizzo M et al. Targeting NRF2, Regulator of Antioxidant System, to Sensitize Glioblastoma Neurosphere Cells to Radiation-Induced Oxidative Stress Oxid Med Cell Longev 2020-06-15 [PMID: 32617133] (WB, Human)
• Rudd SG, Gad H, Sanjiv K et al. MTH1 inhibitor TH588 disturbs mitotic progression and induces mitosis-dependent accumulation of genomic 8-oxodG Cancer Res. 2020-04-20 [PMID: 32312836]
• Moukengue B, Brown HK, Charrier C et al. TH1579, MTH1 inhibitor, delays tumour growth and inhibits metastases development in osteosarcoma model EBioMedicine 2020-03-06 [PMID: 32151797] (Mouse)
• Bialkowski K, Kasprzak KS A profile of 8-oxo-dGTPase activities in the NCI-60 human cancer panel: Meta-analytic insight into the regulation and role of MTH1 (NUDT1) gene expression in carcinogenesis Free Radic. Biol. Med. 2019-12-25 [PMID: 31883466] (WB, Human)
• Wang J, Nagy N, Masucci MG The Epstein-Barr virus nuclear antigen-1 upregulates the cellular antioxidant defense to enable B-cell growth transformation and immortalization Oncogene 2019-09-11 [PMID: 31511648] (WB)
• McPherson LA, Troccoli CI, Ji D et al. Increased MTH1-specific 8-oxodGTPase activity is a hallmark of cancer in colon, lung and pancreatic tissue DNA Repair (Amst.) 2019-07-05 [PMID: 31311767] (WB, Human)
• Zhou W, Ma L, Yang J et al. Potent and specific MTH1 inhibitors targeting gastric cancer Cell Death Dis 2019-06-04 [PMID: 31164636] (WB, Human)
• Pompsch M, Vogel J, Classen F et al. The presumed MTH1-inhibitor TH588 sensitizes colorectal carcinoma cells to ionizing radiation in hypoxia BMC Cancer 2018-11-29 [PMID: 30497423] (WB, Human)
• Zou X, Owusu M, Harris R et al. Validating the concept of mutational signatures with isogenic cell models Nat Commun 2018-05-01 [PMID: 29717121] (WB)
• Fan J, Zhao RQ, Parro C et al. Small molecule inducers of ABCA1 and apoE that act through indirect activation of the LXR pathway. J. Lipid Res. 2018-03-21 [PMID: 29563219] (WB)
• Hashiguchi K, Hayashi M, Sekiguchi M, Umezu K. The roles of human MTH1, MTH2 and MTH3 proteins in maintaining genome stability under oxidative stress. Mutat. Res. 2018-01-23 [PMID: 29482072] (WB, Human)
• Page BDG, Valerie NCK, Wright RHG et al. Targeted NUDT5 inhibitors block hormone signaling in breast cancer cells Nat Commun 2018-01-17 [PMID: 29343827] (WB, Human)
• Eshtad S, Mavajian Z, Rudd SG et al. hMYH and hMTH1 cooperate for survival in mismatch repair defective T-cell acute lymphoblastic leukemia. Oncogenesis 2016-12-05 [PMID: 27918552] (WB, Human)
• Zhou H, Jian Y, Leng Y et al. Human MutT homologue 1 mRNA overexpression correlates to poor response of multiple myeloma. Int J Hematol 2017-03-01 [PMID: 27848179] (Human)
• Huber KV, Olek KM, Muller AC et al. Proteome-wide drug and metabolite interaction mapping by thermal-stability profiling. Nat Methods 2015-11-01 [PMID: 26389571]
• Carter M, Jemth AS, Hagenkort A et al. Crystal structure, biochemical and cellular activities demonstrate separate functions of MTH1 and MTH2. Nat Commun. 2015-08-04 [PMID: 26238318] (WB, Human)
  
  Details:
  MTH1 antibody used for WB on U2OS cells that were transfected with MTH1 siRNA for 96 hours (Figure 4a). The primary antibody was used at a dilution of 1:1,000 with overnight incubation at 4C.
• Giribaldi MG, Munoz A, Halvorsen K et al. MTH1 expression is required for effective transformation by oncogenic HRAS. Oncotarget 2015-05-10 [PMID: 25893378] (Human)
• Shaw D, Wang X, Brown L et al. Infection-derived lipids elicit an immune deficiency circuit in arthropods. Nature Communications. [PMID: 25790189]
• Patel A, Burton DG, Halvorsen K et al. MutT Homolog 1 (MTH1) maintains multiple KRAS-driven pro-malignant pathways. Oncogene 2014-07-14 [PMID: 25023700] (WB, Human)
  
  Details:
  MTH1 antibody used for WB and IHC-P in human species. In WB, the antibody used on lysates of A549, H358, H23 cells treated or not with lentiviral MTH1 shRNA, and several other experiments involving these cell lines. In IHC-P, the antibody used at 1:150 dilution on xenografts created from the transfected cell lines.
• Huber KV, Salah E, Radic B et al. Stereospecific targeting of MTH1 by (S)-crizotinib as an anticancer strategy. Nature 2014-04-10 [PMID: 24695225] (WB, Human)
• Sheng Z, Oka S, Tsuchimoto D et al. 8-Oxoguanine causes neurodegeneration during MUTYH-mediated DNA base excision repair J Clin Invest 2012-12-03 [PMID: 23143307] (ICC/IF, IF/IHC, Mouse)
• Chang IY, Kim JH, Cho KW, Yoon SP. Acute responses of DNA repair proteins and StarD6 in rat hippocampus after domoic acid-induced excitotoxicity Acta Histochem 2012-08-07 [PMID: 22883302] (WB, Mouse)
• Bruel L, Sulzenbacher G, Cervera Tison M et al. alpha-Galactosidase/sucrose kinase (AgaSK), a novel bifunctional enzyme from the human microbiome coupling galactosidase and kinase activities. J Biol Chem 2011-11-25 [PMID: 21931163]
• Rai P, Young JJ, Burton DG et al. Enhanced elimination of oxidized guanine nucleotides inhibits oncogenic RAS-induced DNA damage and premature senescence. Oncogene. 2010-11-15 [PMID: 21076467] (IB, Human)
• Rai et al. PNAS 106 (1):169-174. 2009-01-01 [PMID: 19118192]
• Kennedy, CH et al. Expression of human MutT homologue (hMTH1) protein in primary non-small-cell lung carcinomas and histologically normal surrounding tissue. Free Radic Biol Med;34(11):1447-57. 2003-06-01 [PMID: 12757855] (IHC-Fr, WB, Human)
• Takeshita, F et al. Mechanistic approach of contrasting modifying effects of caffeine on carcinogenesis in the rat colon and mammary gland induced with 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine. Cancer Lett;194(1):25-35. 2003-05-08 [PMID: 12706856] (IHC-P, WB, Rat)

Bioinformatics

• Gene Symbol: NUDT1
• Entrez:
  • 4521(Human)
  • 17766(Mouse)
  • 117260(Rat)
• Uniprot:
  • P36639(Human)