Human Protein Wnt-5a(WNT5A) ELISA kit

Instructions
Code CSB-EL026138HU
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

This Human Protein Wnt-5a (WNT5A) ELISA kit is suitable for qualitatively determining WNT5A concentrations in multiple biological fluids, including in human serum, plasma, and tissue homogenates in vitro. WNT5A is a highly evolutionary conserved non-canonical WNT ligand that activates the non-canonical Wnt pathway. WNT5A signaling plays a critical role in the modulation of normal developmental processes, including proliferation, differentiation, migration, adhesion, and polarity. And it also participates in the regulation of innate immune response and promotes low-grade inflammation. In cancer, it acts as a tumor suppressor or promotes cancer cell invasion, migration, epithelial-mesenchymal transition (EMT), and metastasis.

This kit uses the quantitative sandwich-based enzyme immunoassay technique to measure the amount of human WNT5A in the sample. Standards and samples are respectively added to the microplate wells pre-coated with anti-human WNT5A antibody. Biotin-labeled WNT5A antibody, HRP-avidin, TMB substrate are pipped into the microplate in turn. The capture antibody pre-coated on the plate captures the WNT5A in the rat samples. WNT5A binds to the biotinylated anti-human WNT5A monoclonal antibody. And the biotin on the biotinylated anti-human WNT5A monoclonal antibody binds to the avidin on the enzyme label, forming immune complexes. The color renders blue after the addition of TMB substrate. The addition of the stop solution into the wells immediately turns the blue into yellow. The concentration of WNT5A in the samples is directly proportional to OD (450nm). Each manufactured lot of this ELISA kit was quality tested for criteria such as sensitivity, specificity, and lot-to-lot consistency. See the manual for more information on validation.

Target Name wingless-type MMTV integration site family, member 5A
Alternative Names hWNT 5A ELISA Kit; hWNT5A ELISA Kit; Protein Wnt 5a ELISA Kit; Protein Wnt-5a ELISA Kit; Protein Wnt5a ELISA Kit; Wingless type MMTV integration site family member 5A ELISA Kit; Wnt 5a ELISA Kit; WNT 5A protein ELISA Kit; WNT 5A protein precursor ELISA Kit; WNT5A ELISA Kit; WNT5A protein precursor ELISA Kit; WNT5A_HUMAN ELISA Kit
Abbreviation WNT5A
Uniprot No. P41221
Species Homo sapiens (Human)
Sample Types serum, plasma, tissue homogenates
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 WNT5A 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 % 92  
Range % 90-95  
1:2 Average % 95  
Range % 90-103  
1:4 Average % 84  
Range % 80-92  
1:8 Average % 100  
Range % 98-104  
Recovery
The recovery of human WNT5A 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 83-99  
EDTA plasma (n=4) 97 91-104  
             
             
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 1.839 1.899 1.869 1.685  
5 1.571 1.583 1.577 1.393  
2.5 1.166 1.205 1.186 1.002  
1.25 0.732 0.781 0.757 0.573  
0.625 0.448 0.462 0.455 0.271  
0.312 0.325 0.341 0.333 0.149  
0.156 0.236 0.247 0.242 0.058  
0 0.183 0.185 0.184    
Materials provided
  • A micro ELISA plate ---The 96-well plate has been pre-coated with an anti-human WNT5A 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 WNT5A 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 5-7 working days

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 Q&A
Q:

Does this Elisa kit, CSB-EL026138HU only react with Human cells and not mouse cells?

A:
Thanks for your inquiry! Protein Wnt-5a is protein macromolecule. The protein level AA sequence of different species (human and house) is different, and the structure is also various. Generally it does not make the protein cross-analysis with inter-species. Besides, based on the impact of kit applicability and sample matrix differences and antibody characteristics, we provide this target for different species. Here recommend you to use the corresponding kits for the required species.

Target Background

Function
(From Uniprot)
Ligand for members of the frizzled family of seven transmembrane receptors. Can activate or inhibit canonical Wnt signaling, depending on receptor context. In the presence of FZD4, activates beta-catenin signaling. In the presence of ROR2, inhibits the canonical Wnt pathway by promoting beta-catenin degradation through a GSK3-independent pathway which involves down-regulation of beta-catenin-induced reporter gene expression. Suppression of the canonical pathway allows chondrogenesis to occur and inhibits tumor formation. Stimulates cell migration. Decreases proliferation, migration, invasiveness and clonogenicity of carcinoma cells and may act as a tumor suppressor. Mediates motility of melanoma cells. Required during embryogenesis for extension of the primary anterior-posterior axis and for outgrowth of limbs and the genital tubercle. Inhibits type II collagen expression in chondrocytes.
Gene References into Functions
  1. Impaired Wnt5a signaling is associated with poor placentation and subsequent development of preeclampsia. PMID: 30177057
  2. Decreased hepatic Wnt5a signaling is associated with hepatocellular carcinoma progression and poor prognosis. PMID: 29709351
  3. Annotation of rs11918967 in WNT5A in tissues might be related to obesity. PMID: 28272483
  4. Wnt5a expression is critical for proliferation of RL and VZ progenitors and Purkinje cell dendritogenesis during early embryonic development resulting in retarded development of cerebellum during postnatal stages. PMID: 28205531
  5. Study shows that Wnt5a is upregulated in invasive glioblastoma tissues, and demonstrates that it may regulate the invasion of glioblastoma cells, at least in part via the Daam1/RhoA signaling pathway. PMID: 29207169
  6. The findings suggest that Wnt5a expression may be involved in the inhibition of cell differentiation and the induction of an inflammatory response. PMID: 29286164
  7. High WNT5A expression is associated with gliomas PMID: 28627699
  8. Wnt5a promotes epithelial-to-mesenchymal transition and metastasis in non-small-cell lung cancer, which is involved in the activation of beta-catenin-dependent canonical Wnt signaling. PMID: 29054966
  9. This study suggests that methylation of Wnt pathway genes, in addition to known CpG island methylator phenotype markers, may help predict treatment outcome and survival in patients with CRC[colorectal cancer. PMID: 29869456
  10. non-canonical Wnt5a signalling could play a role in early human trophoblast development by promoting cell proliferation and survival. PMID: 27311852
  11. These findings suggest that WNT-5A modulates fundamental mechanisms that affect airway smooth muscle contraction and thus may be of relevance for airway hyperresponsiveness in asthma. PMID: 27468699
  12. We performed immunohistochemistry for Ki67, p16INK4a, and WNT5A in human HPs ( hyperplastic polyps), sessile serrated adenomas/polyps (SSA/Ps), and traditional serrated adenomas (TSAs) .The distribution of Ki67 and p16INK4a positive cells in TSAs was different from that in HPs and SSA/Ps. PMID: 28627675
  13. Wnt5a-Ror2 signaling enhanced tongue SCC cell aggressiveness and promoted production of MMP-2 following DeltaNp63beta-mediated EMT PMID: 28559016
  14. Here, the authors define WNT5A, a non-canonical Wnt ligand implicated in epithelial differentiation, repair, and cancer, as a direct transcriptional target that is activated by KLF4 in squamous epithelial cells. PMID: 27184424
  15. These findings support that Wnt5a-Ror2 signaling plays a role in UC, support the potential use of Wnt5a as a prognostic marker and provide evidence that Wnt5a signaling may be used as an effective molecular target for novel therapeutic tools. PMID: 28427201
  16. We show that rosiglitazone increases klotho and decreases Wnt5A in tumor cells, reducing the burden of both BRAF inhibitor-sensitive and BRAF inhibitor-resistant tumors in aged, but not young mice. However, when used in combination with PLX4720, tumor burden was reduced in both young and aged mice, even in resistant tumors PMID: 28232477
  17. Serum Wnt5a is elevated and associated with disease severity in heart failure patients. PMID: 28357477
  18. In squamous/adenosquamous carcinoma and adenocarcinoma of the gallbladder positive ROR2 or WNT5a expression is generally associated with a poor prognosis. PMID: 28465645
  19. The Wnt5a and its signaling pathway can regulate fundamental cellular processes, including specification of cell fate, proliferation, and survival. PMID: 28641961
  20. Results show that PTEN and WNT5A expression are directly repressed by miR-26b which promotes colorectal cancer metastasis. PMID: 29160937
  21. The relationship between Wnt5a protein expression and histone H4K20me1 was analyzed. Recruitment of H4K20me1 and SET8 to the Wnt5a promoter and coding regions wa investigated. Results demonstrated that the expression levels of Wnt antagonists were generally low in acute myeloid leukemia (AML), but showed differential expression in acute lymphocytic leukemia (ALL). PMID: 28440495
  22. WNT5A and IL-6 are connected through a positive feedback loop in melanoma cells PMID: 27191257
  23. this study reveals that 14-3-3zeta plays a critical role in Wnt5a/ROR1 signaling, leading to enhanced CLL migration and proliferation. PMID: 28465528
  24. these studies indicate HS1 plays an important role in ROR1-dependent Wnt5a-enhanced chemokine-directed leukemia-cell migration. PMID: 28465529
  25. Study reports that WNT5A bi-directionally regulates epithelial-mesenchymal transition (EMT) in mammary epithelial cells, thereby affecting their migration and invasion. However, the ability of WNT5A to inhibit breast cancer cell migration and invasion is an EMT-independent mechanism that, at least in part, can be explained by decreased CD44 expression. PMID: 27623766
  26. disruption of trans-spliced noncoding RNA RMST expression in human embryonic stem cells results in the upregulation of WNT5A, epithelial-to-mesenchymal transition, and lineage-specific genes/markers. PMID: 27090862
  27. Wnt5a suppressed osteoblastic differentiation through Ror2/JNK signaling in periodontal ligament stem cell-like cells. PMID: 28681925
  28. Genetic blockade of autophagy indicated an unexpected feedback loop whereby knocking down the autophagy factor ATG5 in Wnt5A(high) cells decreased Wnt5A and increased beta-catenin. PMID: 28887323
  29. Low WNT5A expression is associated with prostate cancer. PMID: 28748258
  30. WNT5A signaling regulates 15-PGDH expression. PMID: 27522468
  31. our study showed that, for the first time, different Wnt5a mRNA isoforms play distinct roles in colorectal cancer (CRC) and can be used as novel prognostic markers for CRC in the future. PMID: 28859077
  32. High WNT5a expression is associated with osteoarthritis. PMID: 28777797
  33. We conclude that in RS, WNT5A missense mutations have dominant neomorphic effects that interfere with the function of the wild-type protein. PMID: 28662348
  34. RSPO2 suppresses colorectal cancer metastasis by counteracting the Wnt5a/Fzd7-driven noncanonical Wnt pathway. PMID: 28600110
  35. High expression of WNT5A is associated with squamous cell lung carcinoma. PMID: 27876017
  36. Study shows that WNT5A stimulates dimerization of membrane-anchored FZD4 CRDs and oligomerization of full-length FZD4, which requires the integrity of CRD palmitoleoyl-binding residues. These results suggest that FZD receptors may form signalosomes in response to Wnt binding through the CRDs and that the Wnt palmitoleoyl group is important in promoting these interactions. PMID: 28546512
  37. Taken together with our previous findings, we have replicated our results from the rodent system in a novel human system. We have revealed a unique sequential cascade involving Atg10, Wnt5a, alpha1 integrin, and matrix metalloproteinase-3 in GS/BMP-4-induced differentiation of hiPS cells into odontoblast-like cells at a relatively early stage. PMID: 27639333
  38. Elevated WNT5A expression in obesity may function as a negative regulator of angiogenesis.NEW & NOTEWORTHY Wingless-related integration site 5a (WNT5A) negatively regulates adipose tissue angiogenesis via VEGF-A165b in human obesity. PMID: 28411232
  39. Wnt5a as a master regulator of brain invasion, specifically tumor-promoting stem-like characteristics (TPC), and they provide a therapeutic rationale to target it in patients with glioblastoma. PMID: 28011620
  40. inhibition of WNT-5A in vivo attenuated lung tissue destruction, improved lung function, and restored expression of beta-catenin-driven target genes and alveolar epithelial cell markers in the elastase, as well as in cigarette smoke induced models of COPD. PMID: 27979969
  41. Wnt5A/Ryk signaling might provide novel therapeutic strategies to prevent capillary leakage in systemic inflammation and septic shock. PMID: 27159116
  42. Profound vascular insulin resistance in the visceral adipose tissue arterioles of obese subjects was associated with up-regulated WNT5A-JNK signaling and impaired endothelial eNOS activation. PMID: 27688298
  43. Aberrant activation of WNT pathways in patients with ESRD significantly correlated with vascular calcification. PMID: 27156072
  44. Wnt5A could be a potential therapeutic target for reducing microvascular leakage and edema formation in Th2 driven inflammatory diseases. PMID: 27214384
  45. WNT5A and ROR2 are induced by inflammatory mediators through NF-kB and STAT3 transcription factors, and are involved in the migration of human ovarian cancer cell line SKOV-3. PMID: 28536612
  46. decreased UHRF1 expression is a key initial event in the suppression of DNMT1-mediated DNA methylation and in the consequent induction of senescence via increasing WNT5A expression PMID: 28100769
  47. TrpC5 causes a robust rise in [Ca2+]i, enhanced Wnt5a expression and nuclear translocation of beta-catenin, leading to reduced differentiation and enhanced cancer cell stemness. PMID: 27895148
  48. This study describes the localization and functional role of WNT-5A in human and mouse fibrotic livers. Hepatic WNT-5A expression parallels collagen type I expression. In vivo and in vitro, the myofibroblasts were identified as the key hepatic cells producing WNT-5A. WNT-5A is under control of TGF-beta and its activities are primarily profibrotic. PMID: 28057611
  49. this study shows that Wnt5a promotes differentiation of dendritic cells, but inhibits their maturation PMID: 27641635
  50. we confirmed the requirement of Wnt5a in the deferoxamine-mediated osteoblast-promoting effects by analyzing the matrix mineralization of Wnt5a-deficient cells. The promoting effect of deferoxamine on matrix mineralization in wild-type cells was completely abolished in Wnt5a(-/-) cells. PMID: 27540134

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Involvement in disease Robinow syndrome, autosomal dominant 1 (DRS1)
Subcellular Location Secreted, extracellular space, extracellular matrix
Protein Families Wnt family
Tissue Specificity Expression is increased in differentiated thyroid carcinomas compared to normal thyroid tissue and anaplastic thyroid tumors where expression is low or undetectable. Expression is found in thyrocytes but not in stromal cells (at protein level) (PubMed:157
Database Links

HGNC: 12784

OMIM: 164975

KEGG: hsa:7474

STRING: 9606.ENSP00000264634

UniGene: Hs.643085

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