High Sensitive ELISA Kit for Secreted Frizzled Related Protein 5 (SFRP5) Homo sapiens (Human) High-sensitive ELISA

SARP3; Secreted Apoptosis Related Protein 3; Frizzled-related protein 1b

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  • High Sensitive ELISA Kit for Secreted Frizzled Related Protein 5 (SFRP5) Packages (Simulation)
  • High Sensitive ELISA Kit for Secreted Frizzled Related Protein 5 (SFRP5) Packages (Simulation)
  • High Sensitive ELISA Kit for Secreted Frizzled Related Protein 5 (SFRP5) Results demonstration
  • HEC842Hu.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

Recovery

Matrices listed below were spiked with certain level of recombinant High Sensitive Secreted Frizzled Related Protein 5 (SFRP5) and the recovery rates were calculated by comparing the measured value to the expected amount of High Sensitive Secreted Frizzled Related Protein 5 (SFRP5) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 90-104 94
EDTA plasma(n=5) 96-105 101
heparin plasma(n=5) 98-105 102

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level High Sensitive Secreted Frizzled Related Protein 5 (SFRP5) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level High Sensitive Secreted Frizzled Related Protein 5 (SFRP5) 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 High Sensitive Secreted Frizzled Related Protein 5 (SFRP5) 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) 83-104% 78-102% 85-94% 82-93%
EDTA plasma(n=5) 87-101% 82-102% 81-96% 79-95%
heparin plasma(n=5) 93-101% 84-97% 78-101% 89-98%

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.

High Sensitive ELISA Kit for Secreted Frizzled Related Protein 5 (SFRP5)

Test principle

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to High Sensitive Secreted Frizzled Related Protein 5 (SFRP5). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Sensitive Secreted Frizzled Related Protein 5 (SFRP5). Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain High Sensitive Secreted Frizzled Related Protein 5 (SFRP5), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm ± 10nm. The concentration of High Sensitive Secreted Frizzled Related Protein 5 (SFRP5) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Plasma SFRP5 levels are decreased in Chinese subjects with obesity and type 2 diabetes and negatively correlated with parameters of insulin resistancePubMed: 23290274
  • Sfrp5 correlates with insulin resistance and oxidative stressPubMed: 23398169
  • Balanced duo of anti-inflammatory SFRP5 and proinflammatory WNT5A in childrenPubmed: 24603290
  • SFRP5 correlates with obesity and metabolic syndrome and increases after weight loss in childrenPubmed: 24330025
  • Circulating Sfrp5 is a signature of obesity-related metabolic disorders and is regulated by glucose and liraglutide in humansPubmed: 23185036
  • Low serum level of secreted frizzled-related protein 5, an anti-inflammatory adipokine, is associated with coronary artery diseasePubmed: 24530778
  • Identification of adipokine clusters related to parameters of fat mass, insulin sensitivity and inflammationPubmed:Pmc4072672
  • Activation of noncanonical Wnt signaling through WNT5A in visceral adipose tissue of obese subjects is related to inflammationPubmed:24840810
  • Serum secreted frizzled?related protein 5 levels differentially decrease in patients with hepatitis B virus?associated chronic infection and hepatocellular carcinomaPubmed:25120720
  • Circulating SFRP5 levels are elevated in drug‐na?ve recently diagnosed type 2 diabetic patients as compared with prediabetic subjects and controlsPubmed:25139699
  • The wingless-related integration site-5a/secreted frizzled-related protein-6 system is dysregulated in human sepsisPubmed:25382802
  • Circulating SFRP5 levels are elevated in drug-naPubMed: 25139699
  • Prediction of gestational diabetes mellitus by first trimester serum secreted frizzle-related protein-5 levelsPubMed: 26100762
  • Sfrp5 associates with beta-cell function in humansPubmed:27019073
  • Bone marrow sFRP5 level is negatively associated with bone formation markerspubmed:27986984
  • Plasma Sfrp5 levels correlate with determinants of the metabolic syndrome in Chinese adultspubmed:28303637
  • Inverse associations between serum levels of secreted frizzled-related protein-5 (SFRP5) and multiple cardiometabolic risk factors: KORA F4 studypubmed:28851362
  • 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
  • Longitudinal Increases in Adiposity Contribute to Worsening Adipokine Profile over Time in Mexican AmericansPubmed:29427376
  • Decreased level of the anti-inflammatory adipokines, secreted frizzled-related protein 5 and adiponectin, in high cholesterol diet-induced atherosclerotic ratsarticle-1-352-en.pdf
  • The effect of serum and follicular fluid secreted frizzle-related protein-5 on in vitro fertilization outcomes in patients with polycystic ovary syndromePubmed: 30194556
  • Macrophage LRP1 promotes diet-induced hepatic inflammation and metabolic dysfunction by modulating Wnt signaling
  • A diet high in sugar-sweetened beverage and low in fruits and vegetables is associated with adiposity and a pro-inflammatory adipokine profilePubmed: 30375290
  • Role of wnt5a in Metabolic Inflammation in HumansPubmed: 30137542
  • Association of Adipocytokines With Carotid Intima Media Thickness and Arterial Stiffness in Obstructive Sleep Apnea PatientsPubmed: 32300333
  • Sfrp5/Wnt5a and Leptin/adiponectin levels in the serum and the periarterial adipose tissue of patients with peripheral arterial occlusive diseasePubmed: 33188773
  • 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
  • Secreted Frizzled-Related Protein 5 Ameliorates Vascular Calcification in a Rat Model of Chronic Kidney Disease through the Wnt/β-Catenin Pathway34469882
  • Association of serum uromodulin with adipokines in dependence of type 2 diabetes34920231
  • Association of circulating MR-proADM with all-cause and cardiovascular mortality in the general population: Results from the KORA F4 cohort studyPubmed:34990470
  • Investigating New Sensory Methods Related to Taste Sensitivity, Preferences, and Diet of Mother-Infant Pairs and Their Relationship With Body Composition …Pubmed:35475790
  • Early Pregnancy Serum Concentration of Secreted Frizzled-Related Protein 4, Secreted Frizzled-Related Protein 5, and Chemerin in Obese Women Who Develop …

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