Human Endothelin 1,ET-1 ELISA Kit

Instructions
Code CSB-E07007h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Endothelin 1 (ET-1) ELISA Kit is engineered for accurate measurement of ET-1 levels from samples, including serum, cell culture supernates, urine, and saliva. It uses the Sandwich-ELISA mechanism in combination with the enzyme-substrate chromogenic reaction to measure the ET-1 content in the sample. EDN1 is the most potent endogenous vasoconstrictor generated mainly by the vascular endothelium. It is involved in endothelial dysfunction, inflammation, and vascular remodeling. EDN1 exerts its effects through the interaction with 2 G protein-coupled receptors, the endothelin A (ETA) receptor, and the ETB receptor. Greg Sutton etc. showed that the inflammatory effects of ET-1 and oxLDL on endothelial cells are dependent on an unblocked ETA receptor and functional PKC signaling, ultimately promoting development and progression of atherosclerosis.

The kit has been validated against standards of sensitivity, specificity, precision, linearity, recovery, and lot-to-lot consistency. See more details through the product instructions.

Target Name endothelin 1
Alternative Names ARCND3 ELISA Kit; Big endothelin-1 ELISA Kit; EDN1 ELISA Kit; EDN1_HUMAN ELISA Kit; ET-1 ELISA Kit; ET1 ELISA Kit; HDLCQ7 ELISA Kit; PPET1 ELISA Kit; Preproendothelin 1 ELISA Kit; Preproendothelin-1 ELISA Kit; QME ELISA Kit
Abbreviation EDN1
Uniprot No. P05305
Species Homo sapiens (Human)
Sample Types serum, cell culture supernates, urine, saliva
Detection Range 3.12 pg/mL-200 pg/mL
Sensitivity 0.78 pg/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 ET-1 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 % 89  
Range % 82-97  
1:2 Average % 98  
Range % 94-103  
1:4 Average % 97  
Range % 90-105  
1:8 Average % 87  
Range % 82-91  
Recovery
The recovery of human ET-1 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 87-97  
             
             
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/ml OD1 OD2 Average Corrected  
200 2.722 2.752 2.737 2.628  
100 2.169 2.322 2.246 2.137  
50 1.545 1.528 1.537 1.428  
25 0.882 0.901 0.892 0.783  
12.5 0.551 0.575 0.563 0.454  
6.25 0.341 0.351 0.346 0.237  
3.12 0.245 0.245 0.245 0.136  
0 0.112 0.105 0.109    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human ET-1 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 ET-1 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 high concentration Biotin-antibody to an appropriate working solution.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the high concentration HRP-avidin solution to an appropriate 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 that 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 3-5 working days

Citations

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Target Background

Function
(From Uniprot)
Endothelins are endothelium-derived vasoconstrictor peptides.
Gene References into Functions
  1. The present study demonstrated that the circulating bigET-1 was valuable in the assessment of the severity of CAC. PMID: 29562761
  2. Unfavorable genotype of polymorphic variant of candidate gene participating in endothelial dysfunction EDN1 (Lys198Asn) was associated with changes in levels of their active substances in individuals exposed to mercury. PMID: 30351652
  3. Endothelin-1, nitric oxide, and serotonin are related to blood pressure in adolescent males. PMID: 30271160
  4. this study provides that infants with genotype GT eNOS 894G > T have 3.4-fold higher risk of developing of IVH if they are born before 28 + 6 weeks of gestation. PMID: 28211916
  5. Preeclamptic plasma stimulates the expression of miRNAs in HUVECs, leading to a decrease in endothelin-1 levels. PMID: 29674204
  6. According to the results of the present study, the culture solution with HUVECs affected the differentiation ofdental pulp stem cells (DPSCs). In addition, ET1 may promote the odontoblastic differentiation of DPSCs. PMID: 29845193
  7. EZH2, ET-1, and CD34 may act as biomarkers of aggressive cervical squamous cell carcinoma PMID: 29630085
  8. The plasma D-dimer and ET-1 levels provided a novel marker for treatment selection for the T-segment elevation acute myocardial infarction patients with a type 2 diabetes mellitus history. PMID: 29806659
  9. EDN1 rs5370 polymorphism was associated with large artery stroke development. PMID: 29849817
  10. Endothelin-1 is a regulator of Mena expression in the serous ovarian cancer.Endothelin-1 role in the invadopodian function. PMID: 29439204
  11. Studied effects of dexamethasone on gene expression regulation of sirtuin 1 (SIRT1), interleukin 6 (IL6), and endothelin 1 (EDN1) in gingival derived aging stem cells. Dexamethasone downregulated expression of SIRT1 and IL6 but upregulated EDN1 in stem cells. PMID: 29302812
  12. Baseline big ET-1>0.56pmol/L was independently associated with higher all-cause mortality and HFH among CRT recipients. PMID: 29913121
  13. this study provides the first evidence that a 2-SNP haplotype of the ET-1 gene is associated with increased risk of hormone refractory prostate cancer PMID: 29441862
  14. During exercise, we noted a rapid and significant reduction in venous ET-1 levels in obese subjects but no significant change in lean subjects. PMID: 28977804
  15. Serum levels of miR-31 and endothelin-1are elevated in psoriasis vulgaris and are suggested to play significant roles in the pathophysiology of psoriasis. PMID: 28937251
  16. Plasma big ET-1 level is a valuable independent predictor of ST in patients with coronary stents, especially in the acute coronary syndrome population. PMID: 28942357
  17. Although our study revealed higher serum ET-1 and lower serum alpha-Klotho levels in systemic sclerosis patients compared to healthy controls, there were not any significant correlations between their serum levels with severity of organ involvement. PMID: 28831601
  18. plasma EDN1 levels were significantly increased in Korean patients with fibromyalgia syndrome (FMS) compared with those in healthy controls. EDN1 SNP was revealed to be associated with susceptibility to FMS. PMID: 28901422
  19. miR-200c regulates endothelin-1 induced pulmonary artery smooth muscle cells abnormal proliferation and apoptosis. PMID: 29044995
  20. Increased ET-1 levels after 3 years were positively associated pulse pressure, suggesting subclinical haemodynamic changes with potential premature onset of cardiovascular disease in the black South Africans. PMID: 28726796
  21. The presence of the rs2070699 T allele of EDN1 increased the risk of persistent pulmonary hypertension of the newborn in neonates with respiratory distress. PMID: 27425626
  22. Studied the associations of Protein Z (PROZ G79A) and Endothelin-1 (EDN1 G5665T) polymorphisms with the clinical features of sickle cell disease (SCD) in a cohort of Egyptian children with SCD. PMID: 28548215
  23. Endothelin-1 rs4714384 is an important biomarker of cardiovascular events in end stage renal disease patients. PMID: 28882114
  24. We provided some ways of distinguishing Retinal vein occlusion (RVO)patients in practice and introduced the vasoactivity of the vein itself and the possible involvement of vasoconstrictors such as ET-1 for the pathogenesis of RVO through our clinical studies, some animal experiments, and recent literatures. PMID: 28904960
  25. High levels of plasma endothelin-1 predicted prognosis for patients with hypertrophic cardiomyopathy. PMID: 28587741
  26. highly expressed in the dermal microvascular endothelial cells of hypertrophic scars and keloids PMID: 27892645
  27. ET-1 promoted the proliferation, migration and secretion of extracellular matrix molecules through the protein kinase B (Akt) and extracellular signal-regulated kinase (Erk) signaling pathways in RPE cells in vitro. Thus, ET-1 may play a vital role in the development of PVR. PMID: 28634054
  28. Increased baseline values of ET-1 (and BNP) after acute myocardial infarction, were independently predictive of future cardiovascular events. PMID: 28284500
  29. The EDN1 gene polymorphism may play a significant role in predicting individual's susceptibility toward venous thromboembolism and its clinical progression PMID: 27481876
  30. Epiretinal membrane formation in proliferative diabetic retinopathy involves higher tissue levels of endothelin-1 and fibroblastic activity PMID: 29351334
  31. this study did not find any significant differences of ET-1 level between obstetric antiphospholipid syndrome and thrombotic antiphospholipid syndrome PMID: 27267037
  32. Polymorphic variants of endothelin EDN (K198N) and endothelin receptor type A genes EDN RA (C1222T, C70G, G231A) affected ET plasma concentrations. There was no association between the plasma endothelin levels and the risk factors for normal tension glaucoma. PMID: 27777505
  33. high glucose and free fatty acids reduced NO and increased ET-1 levels induced by acetylcholine through the modulation of intracellular calcium concentrations and endothelial NO synthase phosphorylation, events also reverted by hydroxytyrosol and polyphenol extract. PMID: 25460732
  34. In both the acute and remission phase of thrombotic thrombocytopenic purpura we found increased CT-proET-1 and VWF levels, while Factor H levels decreased compared with healthy controls. In remission, however, the elevated CT-proET-1 levels showed a 22 % decrease when compared with the acute phase. PMID: 26763086
  35. ET-1 increased NSCLC clonal growth, whereas BQ123, ZD4054, lapatinib or gefitinib inhibited proliferation. The results indicate that ET-1 may regulate NSCLC cellular proliferation in an EGFR- and HER2-dependent manner PMID: 28153500
  36. Data indicate that patients with systemic sclerosis (SSc) displayed higher serum levels of VEGF, endothelin-1 and s-Fractalkine, and that the s-Fractalkine levels positively correlated with CD34(+)CD45(-) endothelial progenitor cells (EPC) numbers. PMID: 28320472
  37. Endothelin-1 and its receptors on haemorrhoidal tissue: a potential site for therapeutic intervention PMID: 28095606
  38. we could provide evidence that established stress-related biomarkers ET-1, MCP-1, CGA were differentially regulated among patients with AF compared to healthy controls. PMID: 28886122
  39. Two END1 polymorphisms do not appear to affect the development of hypertension in the Chinese population. PMID: 28692114
  40. TWEAK may stimulate endothelial ET-1 synthesis, through the up-regulation of ECE-1. PMID: 28025386
  41. ET-1 may be a unique predictor of heart failure prognosis PMID: 28398455
  42. The EDNRA SNP rs5335 was independently associated with aneurysms >/=7 mm in size at the time of rupture. Patients with cerebral aneurysms also carrying the G allele of EDNRA SNP rs5335 may develop larger aneurysms before rupture. PMID: 28323184
  43. c-Src tyrosine kinase mediates high glucose-induced endothelin-1 expression PMID: 27102411
  44. This review takes us through the drug discovery journey that led to the discovery, development, and registration of macitentan. [review] PMID: 27534881
  45. Endothelin contributes to the rise in SBP in response to acute hypoxia in patients with severely untreated OSA. PMID: 27906839
  46. Results show a correlation between ET-1 and an impaired liver function, quantified by the maximal liver function capacity test, in critically ill patients. The findings approve the close interaction between TNF-alpha and ET-1 in sepsis. PMID: 28542386
  47. The mean serum levels of endothelin-1 in Egyptian acute myocardial infarction patients was significantly higher than controls. PMID: 27761928
  48. The ratio of serum levels of adiponectin to endothelin strongly correlates with severity of coronary artery disease. PMID: 28781261
  49. Higher levels of circulating endothelin-1 on admission independently predicts subsequent hospital major cardiac events in patients presenting with STEMI. PMID: 27372047
  50. ET-1 is upregulated in giant cell arteritis (GCA) lesions and, by promoting VSMC migration towards the intimal layer, may contribute to intimal hyperplasia and vascular occlusion in GCA. PMID: 28606962

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Involvement in disease Question mark ears, isolated (QME); Auriculocondylar syndrome 3 (ARCND3)
Subcellular Location Secreted
Protein Families Endothelin/sarafotoxin family
Tissue Specificity Expressed in lung, placental stem villi vessels and in cultured placental vascular smooth muscle cells.
Database Links

HGNC: 3176

OMIM: 131240

KEGG: hsa:1906

STRING: 9606.ENSP00000368683

UniGene: Hs.511899

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