Human Telomerase reverse transcriptase(TERT) ELISA kit

Instructions
Code CSB-EL023391HU
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Telomerase reverse transcriptase (TERT) ELISA kit allows for the in vitro quantitative determination of TERT concentrations in serum, plasma, or cell culture supernates. It is not intended for diagnostic use. This assay kit was designed and optimized for cancer research use in humans. The kit has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details.

This assay employs the quantitative sandwich enzyme immunoassay technique, in which TERT in the samples or standards are sandwiched between pre-coated TERT antibody and Biotin-conjugated TERT antibody. HRP-avidin is then added to the wells. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of TERT bound in the initial step. The color development is stopped upon adding the stop solution, and the intensity of the color is measured at 450 nm via a microplate reader. The levels of TERT in the samples can be determined by referring to the O.D. (optical density) of the samples to the standard curve.

TERT is a catalytic subunit of the telomerase. It is involved in a wide range of functions, including the modulation of signal transduction and gene expression, and protection against oxidative damage independent of its telomere elongation activity. TERT also mediates stem cell function, tissue homeostasis, aging, and carcinogenesis.

Target Name telomerase reverse transcriptase
Alternative Names CMM9 ELISA Kit; DKCA2 ELISA Kit; DKCB4 ELISA Kit; EST2 ELISA Kit; HEST2 ELISA Kit; htert ELISA Kit; hTRT ELISA Kit; PFBMFT1 ELISA Kit; TCS1 ELISA Kit; Telomerase associated protein 2 ELISA Kit; Telomerase catalytic subunit ELISA Kit; Telomerase reverse transcriptase ELISA Kit; Telomerase-associated protein 2 ELISA Kit; Telomere Reverse Transcriptase ELISA Kit; TERT ELISA Kit; TERT_HUMAN ELISA Kit; TP2 ELISA Kit; TRT ELISA Kit
Abbreviation TERT
Uniprot No. O14746
Species Homo sapiens (Human)
Sample Types serum, plasma, cell culture supernates
Detection Range 6.25 ng/mL-400 ng/mL
Sensitivity 1.56 ng/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cancer
Assay Principle quantitative
Measurement Sandwich
Precision
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.    
             
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human TERT 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)  
1:1 Average % 94  
Range % 91-101  
1:2 Average % 93  
Range % 89-97  
1:4 Average % 96  
Range % 90-102  
1:8 Average % 93  
Range % 90-96  
Recovery
The recovery of human TERT 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 Type Average % Recovery Range  
Serum (n=5) 92 89-96  
EDTA plasma (n=4) 90 85-96  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
ng/ml OD1 OD2 Average Corrected  
400 2.557 2.657 2.607 2.445  
200 1.824 1.924 1.874 1.712  
100 1.026 1.106 1.066 0.904  
50 0.645 0.655 0.650 0.488  
25 0.370 0.380 0.375 0.213  
12.5 0.261 0.271 0.266 0.104  
6.25 0.205 0.215 0.210 0.048  
0 0.162 0.161 0.162    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human TERT 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 TERT 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-antibody Diluent (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
Materials not provided
  • 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
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 7-14 working days

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Target Data

Function Telomerase is a ribonucleoprotein enzyme essential for the replication of chromosome termini in most eukaryotes. Active in progenitor and cancer cells. Inactive, or very low activity, in normal somatic cells. Catalytic component of the teleromerase holoenzyme complex whose main activity is the elongation of telomeres by acting as a reverse transcriptase that adds simple sequence repeats to chromosome ends by copying a template sequence within the RNA component of the enzyme. Catalyzes the RNA-dependent extension of 3'-chromosomal termini with the 6-nucleotide telomeric repeat unit, 5'-TTAGGG-3'. The catalytic cycle involves primer binding, primer extension and release of product once the template boundary has been reached or nascent product translocation followed by further extension. More active on substrates containing 2 or 3 telomeric repeats. Telomerase activity is regulated by a number of factors including telomerase complex-associated proteins, chaperones and polypeptide modifiers. Modulates Wnt signaling. Plays important roles in aging and antiapoptosis.
Gene References into Functions
  1. Results not only identify quadruplex formation in the first exon promoted by CpG dinucleotide methylation as a regulator of hTERT expression but also provide a possible mechanistic insight into the regulation of gene expression via secondary DNA structures. PMID: 29084850
  2. Findings indicate a critical role for hTERT in regulating the epigenetic clock, in addition to its established role of compensating for cell replication-dependent telomere shortening. PMID: 29374233
  3. CTC1-STN1 terminates TERT while STN1-TEN1 enables C-strand synthesis during telomere replication in colon cancer cells. PMID: 30026550
  4. Results show that TERT promoter (TERTp) mutations were detected in 8.5% of papillary thyroid carcinomas (PTCs) with the C228T mutation being the most frequent. Moreover, three novel TERTp alterations were identified, which may play a role in PTC aggressiveness. Also, TERTp hotspot mutations were highly correlated with BRAF V600E mutation and their coexistence was significantly associated with gender and advanced stage. PMID: 30200646
  5. Presence of TERT promoter mutation is associated with shorter progression free survival and overall survival in meningiomas WHO grade II and III PMID: 29808339
  6. These data suggest that genetic and epigenetic alterations of TERT are associated with TERT upregulation and may predict clinical outcomes in and young adults melanoma. PMID: 28378855
  7. We further characterized those epitopes using enzyme-linked immunosorbent assay, and here we discuss the critical epitope of an anti-TERT mAb, which is applicable for immunohistochemical analysis PMID: 30004263
  8. TERT-p in spitzoid lesions is not necessarily a predictor of poor prognosis PMID: 27930864
  9. there is an association between TERT promoter mutations and MC1R variants in melanoma patients PMID: 27930874
  10. The results indicate that the association with chr5p15.33-Region 2 may be explained by rs36115365, a variant influencing TERT expression via ZNF148 in a manner consistent with elevated TERT in carriers of the C allele. PMID: 28447668
  11. The identified uncommon TERT promoter mutations exacerbate the poor prognosis characteristic of ovarian clear cell carcinoma cases, and the hotspot mutations appear to be a molecular feature of the adenofibroma-associated form of the disease. PMID: 29474637
  12. hTERT contains a BH3-like motif, a short peptide sequence found in BCL-2 family proteins, and interacts with anti-apoptotic BCL-2 family proteins MCL-1 and BCL-xL PMID: 29937479
  13. TERT mutation predicted malignant behavior in three cases of follicular thyroid tumors. PMID: 29860621
  14. the combination of the TERT promoter/BRAFV600E mutations and Ki-67 LI is a promising marker to predict recurrence of PTC. PMID: 28150740
  15. our study verified the correlations of TGFBR2 and hTERT in vitro and suggests that TGFBR2 and hTERT expression may be used as a diagnostic biomarker for cervical dysplasia and carcinoma. PMID: 28195144
  16. epigenetic alterations of the TERT promoter are frequent and associated with advanced disease and poorer clinical outcome in adrenocortical carcinoma. PMID: 29956721
  17. TERT promoter mutations are conserved in the majority of morphologically, spatially and temporally distinct components of a given urothelial carcinoma PMID: 28741734
  18. hTERT Genetic Polymorphisms and Leukocyte Telomere Length Shortenings are associated with Childhood Acute Lymphoblastic Leukemia. PMID: 29936725
  19. TERT Promoter Mutation and Telomere Length is associated with Salivary Gland Tumors. PMID: 28664476
  20. hTERT promoter methylation status revealed non-significant trend towards increase in methylation frequency in head and neck cancer patients compared to healthy individuals. PMID: 30088231
  21. TERT could induce thyroid carcinoma cell proliferation mainly through the PTEN/AKT signaling pathway. PMID: 29901196
  22. There was significant prognostic difference among the 4 glioma subtypes. Combined IDH and TERT gene mutation analysis may be useful for prognostic subgrouping. Notably, IDH1 wild-type cases can be further subdivided into TERT(+ ) or (-) subgroups with significant prognostic difference. PMID: 30220117
  23. EGF significantly upregulated RFPL3 and hTERT protein levels in the nonsmall cell lung cancer cells. RFPL3 and hTERT proteins upregulation by EGF were attenuated by pretreatment with AG1478 and erlotinib. EGF promoted proliferation and inhibited apoptosis; PD98059 decreased RFPL3 and hTERT protein expression; and RFPL3 overexpression increased the expression of hTERT and related MEKpathway proteins PMID: 29749533
  24. TERT protein expression may be regulated by several mechanisms in addition to its promoter mutation. PMID: 29693015
  25. Letter: TERT promoter mutation is a genetic cognate of urothelial papilloma, papillary urothelial neoplasm of low malignant potential and urothelial carcinoma, further supporting the hypothesis that TERT mutation is a frequent and early step in the transformation of many urothelial neoplasms. PMID: 28040359
  26. hTERT promoter mutations associate with aggressive histopathological features, indicating a role in tumour progression in solitary fibrous tumours. PMID: 29703757
  27. Here it was found that there was no statistical difference between human TERT rs2736109 G>A, rs2735940 T>C, rs2853669 A>G, rs2736098 G>A, and rs2736100 T>G polymorphisms that can be associated with risk of Breast Cancer in a Turkish population. PMID: 29506639
  28. the TERT promoter mutation may serve as a biomarker of prognostic stratification in patients with papillary urothelial neoplasm of low malignant potential PMID: 29193225
  29. TERT promoter mutations are very rare in urachal adenocarcinomas (unlike in urothelial carcinoma). PMID: 29047227
  30. Binary logistic regression analysis showed significant association of TERT rs2736100C with gallbladder carcinoma risk. PMID: 29450669
  31. We found that THOR is hypermethylated in pancreatic tumor tissue when compared with normal tissue and that THOR methylation correlates with TERT expression in tumor samples. Our preliminary findings support the diagnostic and prognostic values of THOR in pancreatic cancer. PMID: 29019414
  32. TERT structural rearrangements are associated with metastatic pheochromocytomas. PMID: 28974544
  33. occurrence of TERT promoter mutations has a pivotal role in the disease progression as a secondary genetic event at a time when tumor cells face the need for telomere elongation to allow further proliferation. PMID: 29463038
  34. TERT rs2736098: G>A genotype distribution did not differ significantly between patients with DLBCL and controls. PMID: 28967095
  35. that the rs401681 polymorphism in the TERT-CLPTM1L locus contributes to lung carcinogenesis only in patients harboring an EGFR mutation PMID: 29033187
  36. Mutated Liquid-based FNAs BRAF, N/HRAS and TERT mutations were significantly associated with malignancy regardless of the cytological classification PMID: 29094776
  37. TERT promoter mutations were likely to occur in BRAF V600E positive thyroid cancer. Patients with these 2 combined mutations were more likely to have a poor prognosis and outcome. PMID: 30024548
  38. maternal genetic variations in hTERT may play a contributory role in risk of PTL and PPROM PMID: 29771920
  39. the mutational status of BRAF, NRAS, and TERT promoter genes in 97 melanomas, was investigated. PMID: 29061376
  40. PSMA, TERT, and PDEF may serve as a reference for clinical diagnosis and as potential targets for the malignant tumor of the prostate therapeutics PMID: 28829509
  41. In chronic kidney disease patients, telomerase activity in PBMC was positively correlated with the CKD stage, serum creatinine, potassium and parathormone levels, and negatively correlated with estimated glomerular filtration rate, body mass index, platelet count and serum calcium levels. PMID: 28705647
  42. experimental evidence for association of higher plasma levels of hTERT with overall survival of both low and high grade patients, presenting hTERT as an independent prognostic marker. PMID: 28756592
  43. Tumor suppressor functions of MCPH1/BRIT1 and BRCA1; links with the inactivation of the functional form of hTERT and the activation of dominant negative splice variants of hTERT. PMID: 29860064
  44. results reveal that TERT promoter mutations may contribute significantly to biomarker-based classification of malignant gliomas. PMID: 28868656
  45. Although Genome-Wide Association studies have not been carried out in the field of alcohol-related hepatocellular carcinoma (HCC), common single nucleotide polymorphisms conferring a small increase in the risk of liver cancer risk have been identified. Specific patterns of gene mutations including CTNNB1, TERT, ARID1A and SMARCA2 exist in alcohol-related HCC. [review] PMID: 28296015
  46. Mutation frequency in promoter region in non-acral cutaneous metastatic melanoma. PMID: 27301352
  47. a papillary carcinoma harboring TERT promoter mutation is at higher risk for anaplastic transformation PMID: 28731042
  48. Meta-analysis: The combination of BRAF and TERT promoter mutations could classify PTCs into four distinct risk groups with decreasing aggressiveness as follows: coexisting BRAF and TERT > TERT alone=BRAF alone > no mutations. PMID: 28666074
  49. A total of 13 articles and 15 case-control studies, including 9,157 cases and 11,073 controls, were included in this meta-analysis. Overall, the pooled results indicated that the rs2853669 polymorphism was significantly associated with increased cancer risk in a homozygote comparison model. PMID: 29534075
  50. Authors identified two cancer-specific methylation sites (CpG1 and 2) in the TERT promoter in tumors from GIC patients. Methylated TERT promoter CpG sites 1 and 2 were detectable in patient stool, while only background levels were observed in healthy individuals. PMID: 28754720

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Involvement in disease Aplastic anemia (AA); Dyskeratosis congenita, autosomal dominant, 2 (DKCA2); Pulmonary fibrosis, and/or bone marrow failure, telomere-related, 1 (PFBMFT1); Dyskeratosis congenita, autosomal recessive, 4 (DKCB4); Pulmonary fibrosis, idiopathic (IPF); Melanoma, cutaneous malignant 9 (CMM9)
Subcellular Location Nucleus, nucleolus, Nucleus, nucleoplasm, Nucleus, Chromosome, telomere, Cytoplasm, Nucleus, PML body
Protein Families Reverse transcriptase family, Telomerase subfamily
Tissue Specificity Expressed at a high level in thymocyte subpopulations, at an intermediate level in tonsil T-lymphocytes, and at a low to undetectable level in peripheral blood T-lymphocytes.
Database Links

HGNC: 11730

OMIM: 178500

KEGG: hsa:7015

STRING: 9606.ENSP00000309572

UniGene: Hs.492203

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