Human thiopurine S-methyltransferase (TPMT) ELISA kit
產(chǎn)品詳情
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產(chǎn)品描述：
人硫嘌呤S-甲基轉(zhuǎn)換酶(TPMT)酶聯(lián)免疫試劑盒（CSB-E17858h）為雙抗夾心法ELISA試劑盒，定量檢測血清、血漿、組織勻漿樣本中的TPMT含量。TPMT即硫嘌呤甲基轉(zhuǎn)移酶，是一種參與硫嘌呤類藥物代謝的酶。其基因具有多態(tài)性，不同基因型表達(dá)的酶活性不同。研究機(jī)制主要圍繞其基因多態(tài)性與硫嘌呤類藥物療效及毒副作用的關(guān)聯(lián)，以實(shí)現(xiàn)用藥的個(gè)體化和精準(zhǔn)化。試劑盒檢測范圍為1.56 mU/mL-100 mU/mL，適用于科研領(lǐng)域探究硫嘌呤類藥物代謝機(jī)制、評估患者個(gè)體化用藥方案或開展TPMT遺傳多態(tài)性相關(guān)研究；支持科研人員高效開展藥物代謝動(dòng)力學(xué)、酶活性調(diào)控及疾病機(jī)制等基礎(chǔ)研究。本品僅用于科研，不用于臨床診斷，產(chǎn)品具體參數(shù)及操作步驟詳見產(chǎn)品說明書。
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別名：
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
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縮寫：
TPMT
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Uniprot No.：
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種屬：
Homo sapiens (Human)
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樣本類型：
serum, plasma, tissue homogenates
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檢測范圍：
1.56 mU/mL-100 mU/mL
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靈敏度：
0.39 mU/mL
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反應(yīng)時(shí)間：
1-5h
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樣本體積：
50-100ul
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檢測波長：
450 nm
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研究領(lǐng)域：
Signal Transduction
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測定原理：
quantitative
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測定方法：
Sandwich
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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. | | |
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線性度：
| 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. |
| | Sample | Serum(n=4) | |
| 1:200 | Average % | 87 | |
| Range % | 80-92 | |
| 1:400 | Average % | 98 | |
| Range % | 90-105 | |
| 1:800 | Average % | 100 | |
| Range % | 91-110 | |
| 1:1600 | Average % | 93 | |
| Range % | 86-98 | |
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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. |
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| Sample Type | Average % Recovery | Range | |
| Serum (n=5) | 93 | 85-100 | |
| EDTA plasma (n=4) | 92 | 80-100 | |
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標(biāo)準(zhǔn)曲線：
| These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed. |
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 | mU/ml | OD1 | OD2 | Average | Corrected | | | 100 | 2.239 | 2.152 | 2.196 | 2.044 | | | 50 | 1.854 | 1.760 | 1.807 | 1.655 | | | 25 | 1.311 | 1.294 | 1.303 | 1.151 | | | 12.5 | 0.843 | 0.839 | 0.841 | 0.689 | | | 6.25 | 0.571 | 0.534 | 0.553 | 0.401 | | | 3.12 | 0.380 | 0.359 | 0.370 | 0.218 | | | 1.56 | 0.243 | 0.239 | 0.241 | 0.089 | | | 0 | 0.153 | 0.151 | 0.152 | | |
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本試劑盒所含材料：
- 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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本試劑盒不含材料：
- 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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數(shù)據(jù)處理：
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貨期：
3-5 working days
靶點(diǎn)詳情
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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.
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基因功能參考文獻(xiàn)：
- 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
- In Mexican patients, mutant alleles were detected in 6.2 and 5.2% of systemic lupus erythematosus and rheumatoid arthritis cases, respectively. PMID: 29264794
- TPMT*3C affects TPMT activity in Chinese patients with neuromyelitis optica spectrum disorders. PMID: 29191122
- 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
- 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
- Results show no association between TPMT or COMT single-nucleotide polymorphisms and cisplatin-induced ototoxicity. PMID: 28445188
- 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
- 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
- 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
- This article reviews the thiopurine pharmacogenetics and the methods applied in common practice to evaluate patient's TPMT status. [review] PMID: 28552060
- 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
- TPMT*3C is overrepresented in acute lymphoblastic leukemia (ALL) cases in comparison with non-ALL group PMID: 28476189
- TMPT polymorphisms are associated with 6-mercaptopurine intolerance in children treated for acute lymphoblastic leukemia. PMID: 27577869
- 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
- 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
- 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
- Review/Meta-analysis: thiopurine S-methyltransferase testing for averting drug toxicity. PMID: 27217052
- A point mutation in the thiopurine S-methyltransferase gene that led to exon 5 deletion in the transcribed mRNA. PMID: 26974142
- The aims of this research were to investigate the association of TPMT phenotypes with MDR1 genotypes. PMID: 27453281
- 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
- With respect to TPMT, the variants TPMT*2 and TPMT*3A were not implicated in genetic susceptibility to childhood acute lymphoblastic leukemia PMID: 26845729
- structure-function relationships of TPMT PMID: 26410243
- The TPMT promoter region may serve as a pharmacogenomic biomarker when introducing thiopurine therapy PMID: 26411491
- 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
- 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
- association of TPMT polymorphisms with overall thiopurine-induced adverse drug reactions PMID: 25799415
- Identification of TPMT variants and subsequent dose reduction reduces hematologic events during thiopurine treatment of inflammatory bowel disease. PMID: 26072396
- TPMT*3c mutant allele was associated with azathioprine side effects (leukopenia, alopecia) in Chinese systemic lupus erythematosus patients. PMID: 24322830
- refinements in risk stratification and treatment have reduced the influence of TPMT genotype on treatment outcome in a contemporary protocol. PMID: 25940902
- 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
- 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
- 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
- distribution of the most frequent variants of TPMT gene was similar in a healthy population and patients with inflammatory bowel disease PMID: 23581716
- 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
- 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
- TMPT genotyping appears an important tool to further optimize 6-MP treatment design in Chilean patients with ALL PMID: 24774509
- suggest that germline genetic variation in TPMT and MTHFR do not significantly alter SOS risk in patients exposed to thioguanine PMID: 24737678
- 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
- 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
- Activity measurement performed at diagnosis provides clinicians with information on immediate pharmacokinetic-related adverse events and/or hypermetabolism PMID: 24643197
- 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
- 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
- This study shows the prevalence of TPMT genetic polymorphisms in population of Slovak IBD patients PMID: 23731044
- 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
- In childhood acute lymphoblastic leukaemia, TPMT activity should not be used to predict heterozygosity particularly in blood samples obtained at disease diagnosis PMID: 23252716
- 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
- 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
- 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
- A high frequency and diversity of variant TPMT genotypes was found in this series with predominance of the TPMT*3B allele PMID: 23844534
- 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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亞細(xì)胞定位：
Cytoplasm.
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蛋白家族：
Class I-like SAM-binding methyltransferase superfamily, TPMT family
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數(shù)據(jù)庫鏈接：
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