Human Elastin,ELN ELISA Kit

Instructions
Code CSB-E09338h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Elastin (ELN) ELISA Kit is engineered for accurate measurement of ELN levels from samples, including serum, plasma, cell culture supernates, tissue homogenates, cell lysates, and urine. It uses the Sandwich-ELISA mechanism in combination with the enzyme-substrate chromogenic reaction to measure the ELN content in the sample. ELN is an essential extracellular matrix (ECM) protein of higher vertebrates and the core protein of elastic fibers. As a structural protein, ELN confers elasticity and resilience to various vertebrate tissues and organs such as lungs, skin, and elastic ligaments. In addition to influencing the architecture and biomechanical properties of the extracellular matrix, ELN also plays a vital role in various physiological processes. Some elastin-derived peptides (EDPs) promote angiogenesis and involved in the modulation of various cell activities, including cell adhesion, chemotaxis, migration, proliferation, protease activation, and apoptosis.

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 elastin
Alternative Names Elastin ELISA Kit; ELN ELISA Kit; ELN_HUMAN ELISA Kit; SVAS ELISA Kit; Tropoelastin ELISA Kit; WBS ELISA Kit; WS ELISA Kit
Abbreviation ELN
Uniprot No. P15502
Species Homo sapiens (Human)
Sample Types serum, plasma, cell culture supernates, tissue homogenates, cell lysates, urine
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 Signal Transduction
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 ELN 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)  
1:100 Average % 84  
Range % 80-100  
1:200 Average % 97  
Range % 91-105  
1:400 Average % 97  
Range % 92-112  
1:800 Average % 93  
Range % 86-98  
Recovery
The recovery of human ELN 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) 95 89-100  
EDTA plasma (n=4) 97 90-106  
             
             
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 2.702 2.934 2.818 2.717  
100 2.196 2.184 2.190 2.089  
50 1.497 1.462 1.480 1.379  
25 0.885 0.876 0.881 0.780  
12.5 0.477 0.486 0.482 0.381  
6.25 0.244 0.256 0.250 0.149  
3.12 0.152 0.161 0.157 0.056  
0 0.101 0.100 0.101    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human ELN 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 ELN 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

Customer Reviews and Q&A

 Customer Reviews
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Sample type: Cell culture supernatant

Sample species: Human

Review: The error was not felt between wells of the plate, and it was a reliable kit. The box was easy to use and I liked it.

By Anonymous

Sample type: Biological fluids(urine)

Sample species: Human

Review: I used this kit to determine the elastin concentrations on Human urine samples with no dilution in duplicate and obtained %CV values of 0.2-24.4%. All my values were in the detection range, however, they were in the lower end of the range. Regarding group differences, it is too soon to tell, however, I will be purchasing 10 more kits that will allow me to analyze these differences.

By Anonymous

Target Background

Function
(From Uniprot)
Major structural protein of tissues such as aorta and nuchal ligament, which must expand rapidly and recover completely. Molecular determinant of the late arterial morphogenesis, stabilizing arterial structure by regulating proliferation and organization of vascular smooth muscle (By similarity).
Gene References into Functions
  1. The functional role of PAX5-ELN as a potent oncoprotein in B-cell acute lymphoblastic leukemia development. PMID: 30257940
  2. Elastin plays an important role in the growth of A. fumigatus and that it participates in the formation of thick biofilm. PMID: 29452851
  3. Molecular model of human tropoelastin and implications of associated mutations PMID: 29946030
  4. Study implies that the rs2856728 variant in elastin gene polymorphisms might play crucial roles in the development and pathogenesis of intracranial aneurysms in Korean population. PMID: 29129841
  5. Degradation of tropoelastin and skin elastin by neprilysin PMID: 29196110
  6. exogenous growth factors enhance the expression of cola1, cola3, and Elastin, which is probably regulated via activating MAPK signaling pathway. PMID: 29185160
  7. In conclusion, our results support the view that lysyl oxidase (LOX) and tropoelastin are present on the cell surface and suggests the possibility that lysine oxidation by LOX precedes tropoelastin deposition onto microfibrils. PMID: 28966236
  8. Elastin degradation was correlated with age in COPD patients, smoker controls, and non-smoker controls. The correlation was weaker in the smoker control group compared with the never-smoker control or COPD group. PMID: 27587547
  9. The aim was to examine if the serum concentrations of elastin-related proteins correlate to signs of cardiovascular diseases in patients with Diabetes mellitus type 2. PMID: 28715234
  10. There is evidence that the ELN variant INT20 1315T > C is implicated in the development of intracranial aneurysm. PMID: 27687579
  11. Direct gene sequencing of ELN confirmed the diagnosis showing a previously undescribed c.2156del (p.Gly719Glufs*36) mutation in exon 30 of ELN gene. This mutation results in a shift of the reading frame. PMID: 28383366
  12. Here we report a second adult Williams-Beuren syndrome (WBS)patient with emphysema where the diagnosis of WBS was established subsequent to the discovery of severe bullous emphysema. Haploinsufficiency of ELN likely contributed to this pulmonary manifestation of WBS. PMID: 28574231
  13. the study contributes to a better understanding of the correlation between genotypic and elastin-related phenotypic features of Williams-Beuren syndrome patients PMID: 27311421
  14. We herein report the case of a Japanese female patient presenting with multiple arteriopathy including moyamoya disease, a tortuosity of abdominal arteries and pulmonary hypertension due to peripheral pulmonary artery stenosis. This case suggests the possible progression of cerebral arteriopathy including moyamoya disease in patients with elastin mutations PMID: 27080061
  15. These results indicate that elastin neoepitopes generated by the same proteases but at different amino acid sites provide different tissue-related information depending on the disease in question. PMID: 23805173
  16. Tropoelastin interacts with cells through cell surface receptors including integrins and glycosaminoglycans (GAGs); study mapped a cell-interactive sequence of tropoelastin to domain 17 and the first six amino acids of domain 18. PMID: 28544621
  17. Tropoelastin acts through a PI3K-specific pathway that leads to the phosphorylation of eNOS to enhance nitric oxide production in endothelial cells. PMID: 27175893
  18. Deficient circumferential growth is the predominant mechanism for moderate obstructive aortic disease resulting from partial elastin deficiency in Williams syndrome. PMID: 28254817
  19. In this report we describe a three-generation family suffering from supravalvular aortic stenosis, various other arterial stenoses, sudden death, and intracranial aneurysms. A frameshift mutation in exon 12 of the elastin gene, not described before, was detected in the affected family members. PMID: 27866049
  20. Data show that skin aging is associated with the decomposition of elastin fibers, which is more pronounced in sun-exposed tissue. PMID: 27569260
  21. Transcriptional and posttranscriptional mechanisms contribute to the dysregulation of elastin expression and elastogenesis in Schimke immuno-osseous dysplasia. PMID: 26309238
  22. Neutrophil elastase degraded elastin fragments are increased in the serum of patients with idiopathic pulmonary fibrosis and lung cancer compared to controls. PMID: 25935650
  23. Data suggest that MFAP4 (microfibrillar-associated protein 4) binds tropoelastin, fibrillin-1/2, and elastin cross-linking amino acid desmosine; MFAP4 co-localizes with fibrillin-1-positive fibers; MFAP4 actively promotes tropoelastin self-assembly. PMID: 26601954
  24. In asthma, MMP-1 and MMP-2 mediate IL-13-induced suppression of ELN expression in airway fibroblasts. PMID: 26074138
  25. Current knowledge about elastin peptides relevant to cardiovascular pathologies to further delineate their potential application in cardiovascular disease. [Review] PMID: 25794852
  26. elastin may be important for plaque stability PMID: 25803692
  27. Data suggest that cross-linking involving desmosine and isodesmosine residues in human tropoelastin and bovine elastin contributes to long-term stability of these proteins. PMID: 25604393
  28. Studies indicate that the low sequence complexity, phase behavior, and elastic properties make elastin-like polypeptides (ELPs) an interesting intrinsically disordered proteins (IDP). PMID: 26325592
  29. Supravalvular aortic aneurysms caused by a familial 7q11.23 triplication - including the ELN gene. PMID: 24932728
  30. Up-regulation of elastin and fibulin-5 mRNA levels in ICA were strongly correlated with family history of cardiovascular disease when compared to CCA PMID: 25792650
  31. there may be significantly different associations in genetic variants of elastin between two angiographic phenotypes of polypoidal choroidal vasculopathy (the significant association of rs868005 with Type 2, but not with Type 1) PMID: 25775011
  32. Pathogenetic stimuli induced a significant increase in the expression of LOXL1 and elastic proteins and resulted in their assembly into exfoliation syndrome-like fibrils in vitro. PMID: 25275906
  33. Collectively, these results suggest that dysregulation of LOXL1 expression is a contributing factor to exfoliation disease development. PMID: 25275910
  34. the role of the Asp-72 site in stabilizing the N-terminal segment of human tropoelastin PMID: 25342751
  35. Overall the results indicate that LTBP-2 may have a negative regulatory role during elastic fiber assembly, perhaps in displacing elastin microassemblies from complexes with fibulin-5 and/or cell surface heparan sulfate proteoglycans. PMID: 24148803
  36. The decrease in age-adjusted circulating elastin levels is a potential indicator of eventual cardiovascular dysfunction in medical check-up individuals, as predicted by the risk factors dyslipidemia, hyperuricemia or diabetes. PMID: 23756406
  37. Mutational analysis of the elastin gene identified heterozygosity for a previously unreported de novo c.2184delT mutation in exon 30 not present in either parent. PMID: 24758204
  38. Several extracellular matrix constituents, LOXs, FBN1, and FBLN5, implicated in the development of elastin, are overexpressed in the subepithelial connective tissue extracellular matrix of human pterygium PMID: 25183766
  39. AAA-SMCs have a unique gene expression profile and a proelastolytic phenotype that is augmented by macrophages. PMID: 24080131
  40. Discovery of retinal elastin and its possible role in age-related macular degeneration. PMID: 24232693
  41. Construction and application of elastin like polypeptide containing IL-4 receptor targeting peptide. PMID: 24339977
  42. Tropoelastin modulates TGF-beta1-induced expression of VEGF and CTGF in airway smooth muscle cells. PMID: 23597635
  43. study indicates that the elastin gene may be associated with the formation of IAs, and importantly, that it may also be associated with the rupture of IAs. PMID: 23294012
  44. the sequence of cross-linking domains has a dramatic effect on self-assembly properties of elastin-like polypeptides, and the presence of lysine residues in these domains may serve to prevent inappropriate ordered aggregation. PMID: 24550393
  45. These data reveal a common alphaV integrin-binding theme for tropoelastin: alphaVbeta3 at the C terminus and alphaVbeta5 at the central region of tropoelastin. PMID: 24293364
  46. The authors believe that the Williams-Beuren syndrome chromosome region can be a possible target for further investigation as the genetic basis of keratoconus. PMID: 23167938
  47. Evaluation of these chimeras revealed two exosites contributing to the elastolytic activity of cathepsin V that are distant from the active cleft of the protease and are located in surface loop regions. PMID: 24121514
  48. Elastin binding protein and FKBP65 modulate the kinetics of self-assembly of tropoelastin in an in vitro system. PMID: 24106871
  49. Connective tissue growth factor leads to maximal elastin increase in human umbilical cord-derived mesenchymal stem cells after 7 days of chondroinduction and not in undifferentiated mesenchymal stem cells. PMID: 23542855
  50. Single nucleotide polymorphisms in the IGF2, CCL2, and ELN genes may be associated to the degree and recovery time of non-contact soft tissue injuries. PMID: 23890452

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Involvement in disease Cutis laxa, autosomal dominant, 1 (ADCL1); Supravalvular aortic stenosis (SVAS)
Subcellular Location Secreted, extracellular space, extracellular matrix
Protein Families Elastin family
Tissue Specificity Expressed within the outer myometrial smooth muscle and throughout the arteriolar tree of uterus (at protein level). Also expressed in the large arteries, lung and skin.
Database Links

HGNC: 3327

OMIM: 123700

KEGG: hsa:2006

STRING: 9606.ENSP00000252034

UniGene: Hs.647061

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