ELISA Kit for Wingless Type MMTV Integration Site Family, Member 5A (WNT5A)

Specificity

This assay has high sensitivity and excellent specificity for detection of Wingless Type MMTV Integration Site Family, Member 5A (WNT5A).
No significant cross-reactivity or interference between Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) and the recovery rates were calculated by comparing the measured value to the expected amount of Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 78-97 87
EDTA plasma(n=5) 97-104 101
heparin plasma(n=5) 96-104 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 92-105% 79-98% 93-105% 94-105%
EDTA plasma(n=5) 88-97% 84-101% 91-99% 97-105%
heparin plasma(n=5) 86-95% 96-105% 81-101% 80-96%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Liver International Secreted frizzled-related protein 5 (Sfrp5) decreases hepatic stellate cell activation and liver fibrosis Pubmed:25488180
Journal of Physiology and Biochemistry Wnt5a reverses the inhibitory effect of hyperoxia on transdifferentiation of alveolar epithelial type II cells to type I cells PubMed: 26547443
Liver Secreted frizzled‐related protein 5 (Sfrp5) decreases hepatic stellate cell activation and liver fibrosis PubMed: 25488180
Transl Res Wnt5a is expressed in spondyloarthritis and exerts opposite effects on enthesis and bone in murine organ and cell cultures PubMed: 26163991
不确定杂志名 Untersuchungen der Serum-Konzentrationen des proinflammatorischen Glykoproteins wnt-5a und seines antiinflammatorischen Inhibitors sFRP-5 bei an … servlets:MCRFileNodeServlet
Life Sciences Hepatoprotective effect of hesperidin in hepatocellular carcinoma: Involvement of Wnt signaling pathways. pubmed:28754618
Arteriosclerosis, Thrombosis, and Vascular Biology  Effect of SFRP5 (Secreted Frizzled–Related Protein 5) on the WNT5A (Wingless-Type Family Member 5A)-Induced Endothelial Dysfunction and Its Relevance With … Pubmed:29674475
Stem Cells Reduced Cell Division Control Protein 42 Activity Compromises Hematopoiesis‐Supportive Function of Fanconi Anemia Mesenchymal Stromal Cells Pubmed:29377497
Science Translational Medicine Adipose tissue–derived WNT5A regulates vascular redox signaling in obesity via USP17/RAC1-mediated activation of NADPH oxidases Pubmed: 31534019
J Clin Endocrinol Metab Role of wnt5a in Metabolic Inflammation in Humans Pubmed: 30137542
Cancer Research Host Wnt5a Potentiates Microenvironmental Regulation of Ovarian Cancer Metastasis Pubmed: 31932454
CLINICAL BIOCHEMISTRY Sfrp5/Wnt5a and Leptin/adiponectin levels in the serum and the periarterial adipose tissue of patients with peripheral arterial occlusive disease Pubmed: 33188773
Sao Paulo Medical Journal Association of serum levels of secreted frizzled-related protein 5 and Wnt member 5a with glomerular filtration rate in patients with type 2 diabetes mellitus and … Pubmed: 32491086
Biomaterials An improved osseointegration of metal implants by pitavastatin loaded multilayer films with osteogenic and angiogenic properties 34823885
PLoS One Association of circulating MR-proADM with all-cause and cardiovascular mortality in the general population: Results from the KORA F4 cohort study Pubmed:34990470
J Clin Periodontol Association between bone turnover markers and periodontitis: A population‐based cross‐sectional study Pubmed:35569034
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