Human α-Smooth muscle actin,α-SMA ELISA Kit

Instructions
Code CSB-E09343h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
Have Questions? Leave a Message or Start an on-line Chat

Product Details

Description

The Human α-Smooth muscle actin (α-SMA) ELISA Kit can quantitatively detect the content of α-SMA in human serum, urine, or tissue homogenates. This assay employs the sandwich ELISA mechanism and enzyme-substrate chromogenic reaction to achieve the measurement. The solution develops in proportion to the amount of α-SMA in the sample. And the color intensity can be measured at 450 nm through a microplate reader. The kit has undergone rigorous quality validation in many aspects, including sensitivity, specificity, precision, recovery, linearity, and consistency between batches.

Due to its absence in fibroblasts of normal hearts, α-SMA is the definitive marker for the activated myofibroblasts in hypertrophic and fibrotic hearts and heart failure in humans. α-SMA is also considered as not only a mediator of traction forces in contractile myofibroblasts, but also as a component of the global mechanotransduction system. The study of Arti V.Shinde etc. has shown that α-SMA knockdown attenuates serum-induced contraction of fibroblast-populated collagen pads. And α-SMA overexpression reduces pad contraction, attenuating fibroblast proliferative activity.

Target Name actin, alpha 2, smooth muscle, aorta
Alternative Names a actin ELISA Kit; AAT6 ELISA Kit; ACTA_HUMAN ELISA Kit; ACTA2 ELISA Kit; Actin alpha 2 smooth muscle aorta ELISA Kit; Actin aortic smooth muscle ELISA Kit; Actin; aortic smooth muscle ELISA Kit; ACTSA ELISA Kit; ACTVS ELISA Kit; Alpha 2 actin ELISA Kit; Alpha actin 2 ELISA Kit; Alpha cardiac actin ELISA Kit; alpha sma ELISA Kit; Alpha-actin-2 ELISA Kit; Cell growth inhibiting gene 46 protein ELISA Kit; Cell growth-inhibiting gene 46 protein ELISA Kit; GIG46 ELISA Kit; Growth inhibiting gene 46 ELISA Kit; MYMY5 ELISA Kit
Abbreviation ACTA2
Uniprot No. P62736
Species Homo sapiens (Human)
Sample Types serum, urine, tissue homogenates
Detection Range 3.12 ng/mL-200 ng/mL
Sensitivity 0.78 ng/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 α-SMA 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 % 85  
Range % 80-90  
1:2 Average % 93  
Range % 91-105  
1:4 Average % 94  
Range % 91-110  
1:8 Average % 92  
Range % 83-98  
Recovery
The recovery of human α-SMA 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) 93 87-100  
             
             
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  
200 2.338 2.193 2.266 2.073  
100 1.931 1.893 1.912 1.719  
50 1.547 1.464 1.506 1.313  
25 1.112 1.069 1.091 0.898  
12.5 0.867 0.817 0.842 0.649  
6.25 0.524 0.501 0.513 0.320  
3.12 0.288 0.271 0.280 0.087  
0 0.197 0.188 0.193    
Materials provided
  • A micro ELISA plate --The 96-well plate has been pre-coated with an anti-human α-SMA 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-labled α-SMA 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 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

Customer Reviews and Q&A

 Customer Reviews

There are currently no reviews for this product.

Submit a Review here

Earn $30 Amazon Card or 20μL/μg CUSABIO Trial Size Antibody. Details of rewards >>

 Q&A
Q:

Hello.
I would like to know if CSB-E09343h can be used for detecting alpha SMA in cell culture lysates?

A:
Thanks for your inquiry.
Usually we suggest you test the sample types mentioned in the manual. You can do a pretest if you want to test cell culture lysates.

Target Data

Function Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
Gene References into Functions
  1. Adenocarcinomas showed significantly higher staining scores of both VEGF and alphaSMA than squamous cell carcinomas did. In 42 cases of high CD31 score, five-year survival rate (87%) of patients with lung cancer showing mature tumor vessels was significantly better than that (69%) of patients with immature tumor vessels PMID: 29970531
  2. connective tissue disease including ACTA2 mutations should be considered for aortic dissection in young adult patients. PMID: 28808903
  3. cellular defects due to the ACTA2 mutation in both aortic smooth muscle cells and adventitial fibroblasts may contribute to development of thoracic aortic aneurysms and dissections and proliferative occlusive vascular disease PMID: 28652363
  4. our results seemed to justify the conclusion that ACTA2 did not play a significant role in the pathogenesis of BAV aortopathy. PMID: 29049801
  5. In patients with MYH11 or ACTA2 variants, the effect of intronic variants on splicing was demonstrated on the mRNA level in the induced smooth muscle cell (SMC), allowing classification into pathogenic or nonpathogenic variants. PMID: 28074631
  6. We present a young woman whose ACTA2 mutation was ascertained during pregnancy because of her father's history of dissecting aneurysms. She was delivered at full term by cesarean section and subsequently had severe uterine hemorrhage due to uterine atony. Targeted analysis of the patient's ACTA2 gene revealed she had inherited the N117S variant from her father. PMID: 29202781
  7. During transition from the pluripotent stage towards the neural developmental stage, ACTA2 is differentially expressed in bipolar patient derived cells compared to control derived cells. PMID: 28117838
  8. Two unrelated patients with the heterozygous R189H mutation in ACTA2 and complex congenital heart defects expands the cardiac phenotype of multisystemic smooth muscle dysfunction syndrome. PMID: 28328125
  9. Cells in high glucose for 7 days showed a significant decrease in mRNA expression of CD31 and VE-cadherin, and a significant increase in that of alpha-SMA and collagen I. PMID: 28347704
  10. ACTA2 is the isoform of contractile protein alpha-actin present in vascular smooth muscle cells (SMCs) throughout the arterial tree.4 Pathologic conditions in individuals with ACTA2 mutations show increased deposition of SMCs in the intimal arterial layer, leading to a decreased intraluminal diameter.4 PMID: 28343608
  11. Two novel actin alpha 2 mutations (N117I and L348R) were identified in each familial non-syndromatic thoracic aortic aneurysm proband separately, and an additional novel actin alpha 2 mutation (Y168N) was identified in one patient with sporadic non-syndromatic thoracic aortic aneurysm. PMID: 27431987
  12. The R179H mutation has the potential to affect actin structure and function in both the contractile domain of the cell and the more dynamic cytoskeletal pool of actin, both of which are required for contraction. PMID: 27551047
  13. site-directed mutagenesis revealed several basic amino acid residues in the intermolecular (R267) and intramolecular (K82 and R159) subdomains that are essential for Purbeta transcriptional repressor function in Acta2 promoter-reporter assays. In keeping with their diminished Acta2 repressor activity in fibroblasts, purified Purbeta variants containing an R267A mutation exhibited reduced binding affinity for purine-ric... PMID: 27064749
  14. Data show that the expression profiles of three proteins: E-cadherin, Snail, and alpha-smooth muscle actin were significantly different in extraprostatic extension prostate cancer (PCa) compared with intra-prostatic tumour. PMID: 26701730
  15. ACTA2 mutations are associated with structural disruption and functional impairment of contractile proteins, and predispose to a variety of diffuse vascular diseases including TAAD, CAD, ischemic strokes, and Moyamoya disease. Vascular SMCs are also implicated in vascular remodeling in both physiological and pathological conditions. PMID: 26934405
  16. Parenchymal alphaSMA expression in hepatic tissue appeared to increase only among hepatitis C subjects with fibrosis 3-4. PMID: 26927700
  17. Multisystemic smooth muscle dysfunction syndrome secondary to an ACTA2 mutation. PMID: 26835993
  18. alpha-SMA expression, and MLC phosphorylation, whereas butaprost inhibited TGF-beta2-induced CPCG contraction, actin polymerization, and MLC phosphorylation. PMID: 27082296
  19. Genetic analysis revealed a missense mutation of the ACTA2 gene, encoding for a smooth muscle isoform of alpha-actin. PMID: 27012699
  20. NGF has a role in modulating trkANGFR/p75NTR in alphaSMA-expressing conjunctival fibroblasts from human ocular cicatricial pemphigoid PMID: 26569118
  21. increased level of alphaSMA, a hallmark of epithelial mesenchymal transition in lens epithelial cells, is associated with increased level of histone H4 acetylation at the promoter region of alphaSMA gene(ACTA2) PMID: 25853442
  22. Mutations disrupting p.R179 and p.R258 were associated with significantly increased risk for aortic events, whereas p.R185Q and p.R118Q mutations showed significantly lower risk of aortic events compared with other mutations PMID: 25759435
  23. Alpha-SMA overexpression induced YAP translocation to the nucleus and reduced the high clonogenicity and adipogenic potential of alpha-SMA-negative MSCs. PMID: 26028530
  24. The number and distribution of myofibroblasts and the expression levels of ACTA2 in the fetal membrane may be involved in the mechanisms of development, apoptosis and trophoblastmyofibroblast transformation of the fetal membrane. PMID: 25954927
  25. All the variants analyzed expressed alpha-SMA mildly or moderately, except for the follicular variant that either did not express alpha-SMA or expressed it mildly. PMID: 26470744
  26. alphaSMA was not expressed in benign papillary hyperplastic lesions while it was expressed in papillary carcinoma PMID: 25921136
  27. Transforming growth factor-beta1 induces an up-regulation of alpha-SMA stress fibers in retinal Muller cells and fibroblasts and appears to have a cell-specific effect on intracellular collagen expression. PMID: 26447986
  28. Ca2+- and KCa3.1-dependent processes facilitate "constitutive" alpha smooth muscle actin expression and Smad2/3 signalling in IPF-derived fibroblasts, and thus promote fibroblast to myofibroblast differentiation. PMID: 25476248
  29. The effect of the R258C mutation in SM alpha-actin, is reported. PMID: 26153420
  30. ACTA2, FSP1, and PDGFRa are unfavorable prognostic indicators of esophageal squamous cell carcinoma patients. PMID: 24945657
  31. The immunohistochemical expression of E-cadherin and alpha-smooth muscle actin (alpha-SMA) in 15 cases of pleomorphic adenoma of salivary glands, were investigated. PMID: 25611270
  32. Epithelial-mesenchymal transition-related proteins CK-7 and alpha-SMA colocalized to the intrahepatic biliary epithelial cells in patients with biliary atresia. PMID: 25406900
  33. The ACTA2 gene encodes the smooth muscle alpha2-actin protein. Pathogenic mutations within ACTA2 result in disrupted contractility and are a recognized cause of aortic dissection PMID: 26034244
  34. We report a missense mutation in the smooth muscle alpha-actin (ACTA2; MIM*102620) gene in a 3 generational family from Northern Ireland in which iris flocculi were an ocular marker of the disease. PMID: 24020716
  35. We report a case of ACTA2 mutation in a 3-year-old girl presenting with acute ischemic stroke PMID: 24353327
  36. Women with ACTA2 mutations who are planning to get pregnant should be counseled about this risk of aortic dissection, and proper clinical management should be initiated to reduce this risk. PMID: 24243736
  37. alpha-SMA could be a useful marker for the detection of early stage fibrosis in liver transplant recipients stopping immunosuppressive therapy. PMID: 24966580
  38. This study analyzes the microvascular density (MVD) for CD105+ and alpha-SMA+ vessels and VEGF immunoexpression for 35 oral squamous cell carcinomas and for the associated dysplastic lesions of the lips. PMID: 24715163
  39. This case illustrates the spectrum of systemic malformations that are attributable to mutations in ACTA2 and expands the spectrum of cerebrovascular anomalies that are now known to accompany congenital mydriasis. PMID: 24998021
  40. Identical twin brothers were identified with severe progressive thoracic aortic aneurysm due to ACTA2 mutation. PMID: 25225139
  41. Relationships of alpha-SMA-positive fibroblasts and SDF-1-positive tumor cells with neoangiogenesis in nasopharyngeal carcinoma. PMID: 24877105
  42. High ACTA2 expression is associated with pancreatic cancer. PMID: 25314063
  43. genetic testing revealed new mutations in FBN1, TGFbetaR1, TGFbetaR2 and ACTA2 detected in patients with clinical diagnosis of Marfan Syndrome, Loeys-Dietz Syndrome and Thoracic Aortic Aneurysms and Dissections. PMID: 24793577
  44. Significant down-regulation of the ACTA2 gene, encoding the cytoskeletal protein alpha 2 actin, in response to RHOA knockdown in both osteoblast-like and osteoclast-like cells. PMID: 24840563
  45. Sphingosylphosphorylcholine stimulates alpha-SMA protein expression and human lung fibroblast mediated collagen gel contraction via S1P2 receptor. PMID: 24614064
  46. Suggest dynamic interplay between transcriptional activators Pur-alpha/Pur-beta and repressors in regulating SMalphaA gene output during myofibroblast differentiation. PMID: 24446247
  47. Angiogenesis evaluated through the VEGF and MVD (CD105+ and alpha-SMA+) expression is correlated with the progression and metastasis of gastric cancer and could be considered a prognostic marker of these tumors. PMID: 24322015
  48. ACTA2 regulates c-MET and FAK expression in lung adenocarcinoma cells, which positively and selectively influence metastatic potential. PMID: 23995859
  49. findings confirmed that ACTA2 did not play an important role in the pathogenesis of moyamoya disease PMID: 24024919
  50. Our data suggest that ACTA2 is not a major disease-causing gene for spontaneous cerebral artery dissection PMID: 23879759

Show More

Hide All

Involvement in disease Aortic aneurysm, familial thoracic 6 (AAT6); Moyamoya disease 5 (MYMY5); Multisystemic smooth muscle dysfunction syndrome (MSMDYS)
Subcellular Location Cytoplasm, cytoskeleton
Protein Families Actin family
Database Links

HGNC: 130

OMIM: 102620

KEGG: hsa:59

STRING: 9606.ENSP00000224784

UniGene: Hs.500483

Newsletters

Get all the latest information on Events, Sales and Offers. Sign up for newsletter today.

© 2007-2021 CUSABIO TECHNOLOGY LLC All rights reserved. 鄂ICP备15011166号-1