Human Protein Wnt-3a(WNT3A) ELISA kit

Code CSB-EL026136HU
Size 96T,5×96T,10×96T
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Product Details

Target Name
wingless-type MMTV integration site family, member 3A
Alternative Names
Protein Wnt 3a Precursor ELISA Kit; Protein Wnt-3a ELISA Kit; Wingless type MMTV integration site family member 3A ELISA Kit; Wnt 3a ELISA Kit; wnt3a ELISA Kit; WNT3A protein ELISA Kit; WNT3A_HUMAN ELISA Kit
Abbreviation
WNT3A
Uniprot No.
Species
Homo sapiens (Human)
Sample Types
serum, plasma
Detection Range
0.156 ng/mL-10 ng/mL
Sensitivity
0.039 ng/mL
Assay Time
1-5h
Sample Volume
50-100ul
Detection Wavelength
450 nm
Research Area
Stem Cells
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 WNT3A 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 % 88  
Range % 80-98  
1:2 Average % 95  
Range % 90-100  
1:4 Average % 102  
Range % 91-110  
1:8 Average % 92  
Range % 86-98  
Recovery
The recovery of human WNT3A 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 85-98  
EDTA plasma (n=4) 96 90-103  
             
             
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  
10 2.030 1.952 1.991 1.823  
5 1.549 1.476 1.513 1.345  
2.5 1.092 1.014 1.053 0.885  
1.25 0.712 0.709 0.711 0.543  
0.625 0.487 0.449 0.468 0.300  
0.312 0.372 0.359 0.366 0.198  
0.156 0.273 0.254 0.264 0.096  
0 0.170 0.166 0.168    
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human WNT3A 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 WNT3A 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-conjugateDiluent (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
Storage
Store at 2-8°C. Please refer to protocol.
Lead Time
3-5 working days after you place the order, and it takes another 3-5 days for delivery via DHL or FedEx
Description

This Human WNT3A ELISA Kit was designed for the quantitative measurement of Human WNT3A protein in serum, plasma. It is a Sandwich ELISA kit, its detection range is 0.156 ng/mL-10 ng/mL and the sensitivity is 0.039 ng/mL.

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Target Background

Function
(From Uniprot)
Ligand for members of the frizzled family of seven transmembrane receptors. Functions in the canonical Wnt signaling pathway that results in activation of transcription factors of the TCF/LEF family. Required for normal embryonic mesoderm development and formation of caudal somites. Required for normal morphogenesis of the developing neural tube. Mediates self-renewal of the stem cells at the bottom on intestinal crypts (in vitro).
Gene References into Functions
  1. The miR-766-3p suppressed hepatocellular carcinoma (HCC) cell growth and invasion via a Wnt3a/PRC1 positive regulatory loop, and miR-766-3p may serve as a potential therapeutic target in HCC. PMID: 30145863
  2. High WNT3A expression is associated with colon cancer proliferation and metastasis. PMID: 29948330
  3. Wnt3A regulates the expression of 1,136 genes, of which 662 are upregulated and 474 are downregulated in CCD-18Co cells. A set of genes encoding inhibitors of the Wnt/beta-catenin pathway stand out among those induced by Wnt3A, which suggests that there is a feedback inhibitory mechanism. PMID: 29044515
  4. PCR and western blot results showed that inhibiting the secretion of Wnt3A blocked the Wnt signaling pathway and prevented Nrf2 signaling. Notably, the Wnt inhibitor may serve as a radiosensitizing drug PMID: 28627706
  5. Study found that Wnt3a highly expressed in hepatocellular carcinoma (HCC) tissues, and confirmed that Wnt3a plays an essential role in HCC progression. Activation of the Wnt3a pathway is accompanied by higher expression of Notch3. Furthermore, Wnt3a may be critical for HCC cell cycle and metastasis by regulating cell cycle regulatory proteins and the MAPK pathway. PMID: 28902357
  6. WNT3A supports early stages such as the ALP activity, but it does neither improve later stages of the osteogenic differentiation nor it inhibits the genuine adipogenic differentiation of adipose tissue derived mesenchymal stem cells. PMID: 28549605
  7. SP5 negatively regulates WNT3a transcriptional programs in human pluripotent stem cells. PMID: 29044119
  8. Suggest that GPC5 is able to suppress the lung adenocarcinoma metastasis by competitively binding to Wnt3a and inactivating the Wnt/beta-catenin signaling pathway. PMID: 27806326
  9. WNT3A/beta-catenin signaling inhibition is involved in leflunomide-mediated cytotoxic effects on renal carcinoma cells PMID: 27391060
  10. BRG1 may contribute to colon cancer progression through upregulating WNT3A expression. PMID: 27852072
  11. Here we describe a one-step immobilization technique to covalently bind WNT3A proteins as a basal surface with easy storage and long-lasting activity. We show that this platform is able to maintain adult and embryonic stem cells while also being adaptable for 3D systems PMID: 27411105
  12. study reveals that Foxi1/miR-491-5p/Wnt3a/beta-catenin signaling is critical in the progression of GC. Targeting the pathway described in this study may open up new prospects to restrict the progression of gastric cancer PMID: 28358374
  13. CHIR suppressed the hypertrophic propensity of the MSC-derived cartilage after in vivo implantation to an extent approaching that of WNT3A protein. These results indicate that CHIR may be a promising alternative for WNT3A protein for certain applications of human bone marrow-derived MSCs. PMID: 27633010
  14. PEGylated Wnt3A liposomes associated with skeletal stem cell populations in human bone marrow and promoted osteogenesis. PMID: 28351228
  15. The AChE plays a role in osteoblastic differentiation and is regulated by both Wnt3a and Runx2. PMID: 28607150
  16. In promoting the self-renewal symmetric division of hTERT(high) prostate cancer cells, WNT3a dramatically decreased the ratio of hTERT(high) prostate cancer cells undergoing asymmetric division. Increased WNT/beta-catenin signal activation was also detected in hTERT(high) prostate cancer cells. hTERT-mediated CSC properties were at least partially dependent on beta-catenin. PMID: 28209613
  17. the results indicate neighboring structural elements within full-length Wnt3a affect saposin-like subdomain (SLD) conformational stability. Moreover, SLD function(s) in Wnt proteins appear to have evolved away from those commonly attributed to SAPLIP family members. PMID: 28754322
  18. CPE through its N'-terminal sequence, forms aggregates with Wnt3a and possible endoplasmic reticulum (ER) stress leading to its loss of function. PMID: 27375026
  19. Our findings suggest that Pyk2 plays an important role in the coordination of stabilization of beta-catenin in the crosstalk between Wnt/beta-catenin and Wnt/Ca(2+) signaling pathways upon Wnt3a stimulation in differentiating hNPCs. PMID: 28694190
  20. Macrophage polarization seems to have a key role in the progression of pediatric non-alcoholic fatty liver disease; the modulation of macrophage polarization could drive hepatic progenitor cell response by Wnt3a production and beta-catenin phosphorylation. PMID: 27310371
  21. this study shows that Wnt3a promotes differentiation of dendritic cells, but inhibits their maturation PMID: 27641635
  22. Taken together, our results suggest that canonical Wnt signaling and its antagonist, sFRP1, regulate proliferation of human CSCs. Furthermore, excess sFRP1 in elderly patients causes CSC aging. PMID: 28435069
  23. Thus, our findings provide mechanistic insight into the proposed cross-talk between the Wnt/beta-catenin and Hippo pathways in androgen-independent prostate cancer development. PMID: 28366633
  24. TGF-beta1 signaling might have roles in myofibroblast differentiation in response to orthodontic force. We reveal in vitro that both Wnt3a and TGF-b1 promote myofibroblast differentiation from human periodontal ligament cells PMID: 28223136
  25. Furthermore, pigment epithelium-derived factor (PEDF), a secreted glycoprotein known for its anti-tumor properties, blocked Wnt3a-directed induction of autophagy proteins. Autophagy inhibition was complemented by reciprocal regulation of the oxidative stress enzymes, superoxide dismutase 2 (SOD2) and catalase. PMID: 27557659
  26. we performed a rescue experiment in the JB6 cell line and found that the inhibitory effect of Wnt5a on cell proliferation could be rescued by the addition of Wnt3a. Our data reveal that Wnt5a suppresses the activation of b-catenin signaling during hair follicle regeneration. PMID: 27499692
  27. The level of Wnt3a expression in hepatocellular carcinoma(HCC) patients was obviously higher than that in any group of cases with benign liver diseases. The diagnostic specificity or the area under the receiver operating characteristic curve was 94.34 % or 0.994 in Wnt3a and 69.81 % or 0.710 in AFP for HCC, respectively. PMID: 26577850
  28. Oncogenic Wnt3a expression associated with HBV infection and cirrhotic liver might be an independent prognostic factor for hepatocellular carcinoma. PMID: 27076768
  29. AurkA suppresses the expression of miR-128, inhibitor of wnt3a mRNA stabilization. PMID: 27341528
  30. The pool of ADP-ribosylated Axin, which is degraded under basal conditions, increases immediately following Wnt stimulation in both Drosophila and human cells. PMID: 27138857
  31. The results reveal a novel inhibitory role of Wnt3a on canonical Wnt/beta-catenin signaling and cancer cell proliferation when there is an insufficient blood supply during tumor development. PMID: 26643293
  32. Upregulated Wnt3a in colon cancer cells promoted the capacity to form tube-like structures in the three-dimensional (3-D) culture together with increased expression of endothelial phenotype-associated proteins such as VEGFR2 and VE-cadherin. PMID: 26266404
  33. 72 Wnt target genes higher expressed in triple-negative breast cancer PMID: 25848952
  34. a Wnt-amplified environment was associated with superior pulp healing. PMID: 25556760
  35. Wnt3a can modulate intracellular localisation and secretion of sFRP4. PMID: 25805505
  36. These observations demonstrated the first time that Wnt3a can directly activate MyoD expression through targeting cis-elements in the DE and the L fragment. PMID: 25651906
  37. Wnt-mediated protein stabilization ensures proper mitotic microtubule assembly and chromosome segregation. PMID: 25656539
  38. Wnt3a expression was associated with epithelial mesenchymal transition and promoted colon cancer progression. PMID: 25499541
  39. Wnt3a and Notch3 may promote the metastasis of non-small cell lung cancer and Notch3 upregulation is required for the Wnt3a mediated increased metastatic abilities of non-small cell lung cancer. PMID: 25572698
  40. SNP rs752107 in WNT3A was strongly associated with decreased total hip BMD showing the highest significance under the recessive model PMID: 24584697
  41. miR-195 is key in regulating cell proliferation, cell cycle and apoptosis through targeting Wnt3a. PMID: 25174704
  42. WNT3A alone was a poor inducer of mesoderm from hESCs in defined medium. WNT3A and BMP4 synergized to accelerate mesoderm formation. Added later, WNT3A blocked hematopoiesis and generated mesenchymal colonies. PMID: 24052942
  43. High WNT3A expression is associated with glioblastoma. PMID: 25301448
  44. WNT3A gene expression is associated with isolated Hirschsprung disease polymorphism and disease status. PMID: 24817932
  45. High WNT3A expression is associated with bone destruction in lung and breast cancer. PMID: 25359619
  46. High Wnt3A expression is associated with metastasis in triple negative breast cancer. PMID: 24209998
  47. Application of recombinant Wnt3a showed an anti-proliferative effect on keratinocytes in a dose-dependent manner. PMID: 24686518
  48. these results show that TAZ mediates Wnt3a-stimulated osteogenic differentiation through PP1A, suggesting that the Wnt signal regulates the Hippo pathway. PMID: 24510127
  49. The LRP6 tyrosine mutant increased in signaling activation in response to Wnt3a/beta-catenin stimulation. PMID: 25143377
  50. p66(Shc) plays a vital part in canonical Wnt signaling in the endothelium and mediates Wnt3a-stimulated endothelial oxidative stress and dysfunction. PMID: 25147340

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Subcellular Location
Secreted, extracellular space, extracellular matrix. Secreted.
Protein Families
Wnt family
Tissue Specificity
Moderately expressed in placenta and at low levels in adult lung, spleen, and prostate.
Database Links

HGNC: 15983

OMIM: 606359

KEGG: hsa:89780

STRING: 9606.ENSP00000284523

UniGene: Hs.336930

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