Multiplex Assay Kit for Secreted Frizzled Related Protein 5 (SFRP5) ,etc. by FLIA (Flow Luminescence Immunoassay)

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

(Note: Up to 8-plex in one testing reaction)

Specificity

This assay has high sensitivity and excellent specificity for detection of Secreted Frizzled Related Protein 5 (SFRP5) ,etc. by FLIA (Flow Luminescence Immunoassay).
No significant cross-reactivity or interference between Secreted Frizzled Related Protein 5 (SFRP5) ,etc. by FLIA (Flow Luminescence Immunoassay) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 89-103 95
EDTA plasma(n=5) 83-96 89
heparin plasma(n=5) 84-91 87
sodium citrate plasma(n=5) 81-93 85

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Secreted Frizzled Related Protein 5 (SFRP5) ,etc. by FLIA (Flow Luminescence Immunoassay) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Secreted Frizzled Related Protein 5 (SFRP5) ,etc. by FLIA (Flow Luminescence Immunoassay) 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 Secreted Frizzled Related Protein 5 (SFRP5) ,etc. by FLIA (Flow Luminescence Immunoassay) 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) 87-104% 82-89% 91-98% 78-95%
EDTA plasma(n=5) 81-90% 82-103% 91-105% 90-97%
heparin plasma(n=5) 85-92% 98-105% 78-103% 85-101%
sodium citrate plasma(n=5) 94-103% 92-101% 94-104% 84-98%

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
96-well plate 1 Plate sealer for 96 wells 4
Pre-Mixed Standard 2 Standard Diluent 1×20mL
Pre-Mixed Magnetic beads (22#:SFRP5) 1 Analysis buffer 1×20mL
Pre-Mixed Detection Reagent A 1×120μL Assay Diluent A 1×12mL
Detection Reagent B (PE-SA) 1×120μL Assay Diluent B 1×12mL
Sheath Fluid 1×10mL Wash Buffer (30 × concentrate) 1×20mL
Instruction manual 1

Assay procedure summary

1. Preparation of standards, reagents and samples before the experiment;
2. Add 100μL standard or sample to each well,
    add 10μL magnetic beads, and incubate 90min at 37°C on shaker;
3. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60min at 37°C on shaker;
4. Wash plate on magnetic frame for three times;
5. Add 100μL prepared Detection Reagent B, and incubate 30 min at 37°C on shaker;
6. Wash plate on magnetic frame for three times;
7. Add 100μL sheath solution, swirl for 2 minutes, read on the machine.

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Magazine Citations
Diabetes Research and Clinical Practice Plasma SFRP5 levels are decreased in Chinese subjects with obesity and type 2 diabetes and negatively correlated with parameters of insulin resistance PubMed: 23290274
European Journal of Clinical Investigation Sfrp5 correlates with insulin resistance and oxidative stress PubMed: 23398169
Pediatric Research Balanced duo of anti-inflammatory SFRP5 and proinflammatory WNT5A in children Pubmed: 24603290
Clin Endocrinol (Oxf). SFRP5 correlates with obesity and metabolic syndrome and increases after weight loss in children Pubmed: 24330025
The Journal of Clinical Endocrinology & Metabolism Circulating Sfrp5 is a signature of obesity-related metabolic disorders and is regulated by glucose and liraglutide in humans Pubmed: 23185036
Atherosclerosis Low serum level of secreted frizzled-related protein 5, an anti-inflammatory adipokine, is associated with coronary artery disease Pubmed: 24530778
PLoS One. Identification of adipokine clusters related to parameters of fat mass, insulin sensitivity and inflammation Pubmed:Pmc4072672
J Clin Endocrinol Metab. Activation of noncanonical Wnt signaling through WNT5A in visceral adipose tissue of obese subjects is related to inflammation Pubmed:24840810
Oncology Letters Serum secreted frizzled?related protein 5 levels differentially decrease in patients with hepatitis B virus?associated chronic infection and hepatocellular carcinoma Pubmed:25120720
Diabetes Metab Res Rev. Circulating SFRP5 levels are elevated in drug‐na?ve recently diagnosed type 2 diabetic patients as compared with prediabetic subjects and controls Pubmed:25139699
Clinical &amp; Experimental Immunology The wingless-related integration site-5a/secreted frizzled-related protein-6 system is dysregulated in human sepsis Pubmed:25382802
Diabetes Metab Res Rev Circulating SFRP5 levels are elevated in drug-na PubMed: 25139699
J Matern Fetal Neonatal Med Prediction of gestational diabetes mellitus by first trimester serum secreted frizzle-related protein-5 levels PubMed: 26100762
Eur J Clin Invest Sfrp5 associates with beta-cell function in humans Pubmed:27019073
Osteoporosis International Bone marrow sFRP5 level is negatively associated with bone formation markers pubmed:27986984
Diabetes/Metabolism Research and Reviews Plasma Sfrp5 levels correlate with determinants of the metabolic syndrome in Chinese adults pubmed:28303637
Cardiovascular Diabetology Inverse associations between serum levels of secreted frizzled-related protein-5 (SFRP5) and multiple cardiometabolic risk factors: KORA F4 study pubmed:28851362
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
Obesity Longitudinal Increases in Adiposity Contribute to Worsening Adipokine Profile over Time in Mexican Americans Pubmed:29427376
J Bas Res Med Sci Decreased level of the anti-inflammatory adipokines, secreted frizzled-related protein 5 and adiponectin, in high cholesterol diet-induced atherosclerotic rats article-1-352-en.pdf
Molecular Biology Reports The effect of serum and follicular fluid secreted frizzle-related protein-5 on in vitro fertilization outcomes in patients with polycystic ovary syndrome Pubmed: 30194556
Macrophage LRP1 promotes diet-induced hepatic inflammation and metabolic dysfunction by modulating Wnt signaling
British Journal Of Nutrition A diet high in sugar-sweetened beverage and low in fruits and vegetables is associated with adiposity and a pro-inflammatory adipokine profile Pubmed: 30375290
J Clin Endocrinol Metab Role of wnt5a in Metabolic Inflammation in Humans Pubmed: 30137542
Frontiers in Endocrinology Association of Adipocytokines With Carotid Intima Media Thickness and Arterial Stiffness in Obstructive Sleep Apnea Patients Pubmed: 32300333
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
Kidney Blood Press Res Secreted Frizzled-Related Protein 5 Ameliorates Vascular Calcification in a Rat Model of Chronic Kidney Disease through the Wnt/β-Catenin Pathway 34469882
Cytokine Association of serum uromodulin with adipokines in dependence of type 2 diabetes 34920231
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
JMIR Res Protoc Investigating New Sensory Methods Related to Taste Sensitivity, Preferences, and Diet of Mother-Infant Pairs and Their Relationship With Body Composition … Pubmed:35475790
Diabetology 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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