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  4. Human thiopurine S-methyltransferase (TPMT) ELISA kit

Human thiopurine S-methyltransferase (TPMT) ELISA kit

  • 中文名称：
    人硫嘌呤S-甲基转换酶(TPMT)酶联免疫试剂盒
  • 货号：
    CSB-E17858h
  • 规格：
    96T/48T
  • 价格：
    ￥3600/￥2500
  • 其他：

产品详情

  • 产品描述：

    This Human TPMT ELISA Kit was designed for the quantitative measurement of Human TPMT protein in serum, plasma, tissue homogenates. It is a Sandwich ELISA kit, its detection range is 1.56 mU/mL-100 mU/mL and the sensitivity is 0.39 mU/mL.

  • 别名：
    HGNC:12014 ELISA Kit; S adenosyl L methionine thiopurine S methyltransferase ELISA Kit; Thiopurine methyltransferase ELISA Kit; Thiopurine S methyltransferase ELISA Kit; Thiopurine S-methyltransferase ELISA Kit; TPMT ELISA Kit; TPMT_HUMAN ELISA Kit
  • 缩写：
    TPMT
  • Uniprot No.：
  • 种属：
    Homo sapiens (Human)
  • 样本类型：
    serum, plasma, tissue homogenates
  • 检测范围：
    1.56 mU/mL-100 mU/mL
  • 灵敏度：
    0.39 mU/mL
  • 反应时间：
    1-5h
  • 样本体积：
    50-100ul
  • 检测波长：
    450 nm
  • 研究领域：
    Signal Transduction
  • 测定原理：
    quantitative
  • 测定方法：
    Sandwich
  • 精密度：
    Intra-assay Precision (Precision within an assay): CV%<8%
    Three samples of known concentration were tested twenty times on one plate to assess.
    Inter-assay Precision (Precision between assays): CV%<10%
    Three samples of known concentration were tested in twenty assays to assess.
  • 线性度：
    To assess the linearity of the assay, samples were spiked with high concentrations of human TPMT in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
    　SampleSerum(n=4)
    1:200Average %87
    Range %80-92
    1:400Average %98
    Range %90-105
    1:800Average %100
    Range %91-110
    1:1600Average %93
    Range %86-98
  • 回收率：
    The recovery of human TPMT spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
    Sample TypeAverage % RecoveryRange
    Serum (n=5) 9385-100
    EDTA plasma (n=4)9280-100
  • 标准曲线：
    These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
    mU/mlOD1OD2AverageCorrected
    1002.239 2.152 2.196 2.044
    501.854 1.760 1.807 1.655
    251.311 1.294 1.303 1.151
    12.50.843 0.839 0.841 0.689
    6.250.571 0.534 0.553 0.401
    3.120.380 0.359 0.370 0.218
    1.560.243 0.239 0.241 0.089
    00.153 0.151 0.152 　
  • 本试剂盒所含材料：
    • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human TPMT antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
    • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
    • One vial Biotin-labeled TPMT antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
    • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
    • One vial Biotin-antibodyDiluent (15 ml/bottle) ---Dilute the Biotin-antibody.
    • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
    • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
    • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
    • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
    • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
    • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
    • An instruction manual

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  • 本试剂盒不含材料：
    • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
    • An incubator can provide stable incubation conditions up to 37°C±5°C.
    • Centrifuge
    • Vortex
    • Squirt bottle, manifold dispenser, or automated microplate washer
    • Absorbent paper for blotting the microtiter plate
    • 50-300ul multi-channel micropipette
    • Pipette tips
    • Single-channel micropipette with different ranges
    • 100ml and 500ml graduated cylinders
    • Deionized or distilled water
    • Timer
    • Test tubes for dilution

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  • 数据处理：
  • 货期：
    3-5 working days

产品评价

靶点详情

  • 功能：
    Catalyzes the S-methylation of thiopurine drugs such as 6-mercaptopurine (also called mercaptopurine, 6-MP or its brand name Purinethol) and 6-thioguanine (also called tioguanine or 6-TG) using S-adenosyl-L-methionine as the methyl donor. TPMT activity modulates the cytotoxic effects of thiopurine prodrugs. A natural substrate for this enzyme has yet to be identified.
  • 基因功能参考文献：
    1. Mutation rate of NUDT15 in Chinese inflammatory bowel disease patients is higher than that of TPMT. NUDT15 polymorphism is a better predictor for AZA-induced leukopenia than TPMT polymorphism. PMID: 29491687
    2. In Mexican patients, mutant alleles were detected in 6.2 and 5.2% of systemic lupus erythematosus and rheumatoid arthritis cases, respectively. PMID: 29264794
    3. TPMT*3C affects TPMT activity in Chinese patients with neuromyelitis optica spectrum disorders. PMID: 29191122
    4. Patients also developed infection who mostly (71%) needed hospitalization. None of the studied G238C, G460A and A719G TPMT variant alleles were detected. Infections and febrile neutropenia were common causes of 6-PM dose modification and interruption. PMID: 29321348
    5. ANCA associated vasculitis (AAV) patients treated with cyclophosphamide induction and azathioprine maintenance therapy were included and followed for 60 months. TPMT genotype and activity were not independent predictors of relapse, and could not predict leukopenia or other adverse effects from azathioprine. PMID: 29630648
    6. Results show no association between TPMT or COMT single-nucleotide polymorphisms and cisplatin-induced ototoxicity. PMID: 28445188
    7. Of the 14 patients intolerant to thiopurines, NGS identified deleterious TPMT variants in 5 individuals whereas the biochemical test identified 8 individuals as intolerant (sensitivity 35.7% and 57.14%; specificity 93.75% and 50% respectively). SKAT-O test identified a significant association between MOCOS gene and TPMT activity (p = 0.0015), not previously reported. PMID: 27703193
    8. A statistically significant correlation between the presence of the G460A mutation and the development of leucopenia after the administration of thiopurines was observed. PMID: 28857898
    9. genetic association studies in cohort in Denmark: Data suggest that standard immunosuppression treatment failure in patients with autoimmune hepatitis is not associated with thiopurine S-methyltransferase genetic variants or HLA-DRB1*03 genotype in the population studied. (HLA-DRB1 = major histocompatibility complex class II DR beta 1) PMID: 28374975
    10. This article reviews the thiopurine pharmacogenetics and the methods applied in common practice to evaluate patient's TPMT status. [review] PMID: 28552060
    11. study aimed to determine the relationship between thiopurine metabolite levels and therapeutic response, and to investigate the association of NUDT15, TPMT, and thiopurine metabolites with leukopenia in patients with CD PMID: 29206869
    12. TPMT*3C is overrepresented in acute lymphoblastic leukemia (ALL) cases in comparison with non-ALL group PMID: 28476189
    13. TMPT polymorphisms are associated with 6-mercaptopurine intolerance in children treated for acute lymphoblastic leukemia. PMID: 27577869
    14. The frequency of TPMT mutation in a homogeneous Sicilian cohort of patients with inflammatory bowel disease, autoimmune and hematological disorders was low. TPMT genotyping is not a sensitive tool for predicting thiopurine-induced leukopenia. PMID: 27665263
    15. meta-analysis of three independent GWAS of TPMT activity; only genetic variants mapping to chromosome 6, including the TPMT gene region, were significantly associated with TPMT activity;finding is consistent with TPMT genotype being the primary determinant of TPMT activity PMID: 27770449
    16. In a GWAS, TPMT activity in patients behaves as a monogenic trait, further bolstering the utility of TPMT genetic testing in the clinic PMID: 27564568
    17. Review/Meta-analysis: thiopurine S-methyltransferase testing for averting drug toxicity. PMID: 27217052
    18. A point mutation in the thiopurine S-methyltransferase gene that led to exon 5 deletion in the transcribed mRNA. PMID: 26974142
    19. The aims of this research were to investigate the association of TPMT phenotypes with MDR1 genotypes. PMID: 27453281
    20. The aim of the study was to investigate frequencies of TPMT and ITPA polymorphisms in Lithuanian inflammatory bowel disease patients and analyze their association with azathioprine-related adverse events. PMID: 26674571
    21. With respect to TPMT, the variants TPMT*2 and TPMT*3A were not implicated in genetic susceptibility to childhood acute lymphoblastic leukemia PMID: 26845729
    22. structure-function relationships of TPMT PMID: 26410243
    23. The TPMT promoter region may serve as a pharmacogenomic biomarker when introducing thiopurine therapy PMID: 26411491
    24. The most frequently occurring nonfunctional TPMT allele in Croatian population is TPMT*3A. Variant genotypes were statistically significantly more common in Crohn's disease subgroup than in ulcerative colitis subgroup. PMID: 27333713
    25. Association of TPMT polymorphisms with overall azathioprine-induced adverse drug reactions, bone marrow toxicity and gastric intolerance, but not with hepatotoxicity (meta-analysis). PMID: 26633017
    26. association of TPMT polymorphisms with overall thiopurine-induced adverse drug reactions PMID: 25799415
    27. Identification of TPMT variants and subsequent dose reduction reduces hematologic events during thiopurine treatment of inflammatory bowel disease. PMID: 26072396
    28. TPMT*3c mutant allele was associated with azathioprine side effects (leukopenia, alopecia) in Chinese systemic lupus erythematosus patients. PMID: 24322830
    29. refinements in risk stratification and treatment have reduced the influence of TPMT genotype on treatment outcome in a contemporary protocol. PMID: 25940902
    30. We report on the presence of the TPMT*3C and *3A mutant alleles in the Libyan population. Therefore, monitoring the patients to be treated with doses of thiopurine drugs for TPMT variants is worthwhile to avoid the development of severe myelosuppression. PMID: 25819542
    31. this paper shows that the influence of TPMT and COMT on the development of cisplatin-induced hearing loss may be less important than previously suggested. PMID: 25551397
    32. Results show complete sequence-based screening study evaluating all TPMT variants in Asian populations some of them may ne of relevance in Korean population. PMID: 25564374
    33. distribution of the most frequent variants of TPMT gene was similar in a healthy population and patients with inflammatory bowel disease PMID: 23581716
    34. The prevalence of TPMT genotypes was high among Brazilian patients. Variants genes 2 and 3C may be associated with azathioprine pancreatic toxicity in a IBD southeastern Brazilian population PMID: 24696613
    35. Data suggest risk of myelotoxicity of high-dose methotrexate during methotrexate/6-mercaptopurine maintenance chemotherapy of childhood acute leukemia is not associated with heterozygosity in thiopurine methyltransferase in Nordic population studied. PMID: 25347948
    36. TMPT genotyping appears an important tool to further optimize 6-MP treatment design in Chilean patients with ALL PMID: 24774509
    37. suggest that germline genetic variation in TPMT and MTHFR do not significantly alter SOS risk in patients exposed to thioguanine PMID: 24737678
    38. TPMT*37 introduces a premature stop codon at position 216, resulting in loss of the last 29 amino acid residues from the C terminal of the TPMT protein. PMID: 24710034
    39. plays a key role in the metabolism of azathioprine/6- mercaptopurine (6-MP). Mutations in the enzyme lead to generation of excess thioguanine, which causes neutropenia via suppression of neutrophils. PMID: 23996738
    40. Activity measurement performed at diagnosis provides clinicians with information on immediate pharmacokinetic-related adverse events and/or hypermetabolism PMID: 24643197
    41. Heart transplantation recipients with TPMT genetic variant alleles who are treated with AZA develop acute rejection earlier, more frequently, and of greater severity. PMID: 24121523
    42. The study focused on explaining how a locally occurred TPMT A167G substitution propagated through hydrogen bonds alteration to induce structural modification which affects both thiopurine and S-adenosylmethionine receptors. PMID: 23025308
    43. This study shows the prevalence of TPMT genetic polymorphisms in population of Slovak IBD patients PMID: 23731044
    44. TPMT*3A and the TPMT*3C, which cause the largest decrease in enzyme activity, were both variant alleles detected in the Tunisian population PMID: 23553048
    45. In childhood acute lymphoblastic leukaemia, TPMT activity should not be used to predict heterozygosity particularly in blood samples obtained at disease diagnosis PMID: 23252716
    46. differential contribution of the enzyme TPMT to the cytotoxicity of the two thiopurines is probably due to its role in formation of the meTIMP, the cytotoxic methylated metabolite of 6-MP PMID: 23811272
    47. A predictive model combining variants in TPMT, ABCC3, and COMT with clinical variables significantly improved the prediction of hearing-loss development as compared with using clinical risk factors alone PMID: 23588304
    48. our results indicated that TPMT or COMT genetic variation was not related to cisplatin ototoxicity in children with cancer and did not influence cisplatin-induced hearing damage in laboratory models PMID: 23820299
    49. A high frequency and diversity of variant TPMT genotypes was found in this series with predominance of the TPMT*3B allele PMID: 23844534
    50. In a study of Italian myasthenia gravis patients treated with azathioprine, a new TPMT haplotype, TPMT* 3E, was observed only in association with intolerance. PMID: 23400745

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  • 亚细胞定位：
    Cytoplasm.
  • 蛋白家族：
    Class I-like SAM-binding methyltransferase superfamily, TPMT family
  • 数据库链接：

    HGNC: 12014

    OMIM: 187680

    KEGG: hsa:7172

    STRING: 9606.ENSP00000312304

    UniGene: Hs.444319