Multiplex Assay Kit for Secreted Frizzled Related Protein 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array)

FRP-4; FRPHE; Frizzled protein, human endometrium

Specificity

This assay has high sensitivity and excellent specificity for detection of Secreted Frizzled Related Protein 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array).
No significant cross-reactivity or interference between Secreted Frizzled Related Protein 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Secreted Frizzled Related Protein 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array) and the recovery rates were calculated by comparing the measured value to the expected amount of Secreted Frizzled Related Protein 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 82-94 89
EDTA plasma(n=5) 99-105 102
heparin plasma(n=5) 79-91 86

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Secreted Frizzled Related Protein 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array) 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 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array) 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 4 (SFRP4) ,etc. by CBA (Cytometric Bead Array) 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) 90-103% 90-97% 99-105% 98-105%
EDTA plasma(n=5) 78-96% 80-92% 81-104% 94-101%
heparin plasma(n=5) 84-103% 98-105% 92-105% 84-104%

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

Assay procedure summary

1. Add 200μL analysis buffer solution to each well of the plate for pre-wetting;
2. Preparation of standards, reagents and samples before the experiment;
3. Add 100μL standard or sample to each well, add 10μL magnetic beads, and incubate 120 minutes protect from light at 25°C on shaker;
4. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent A. Incubate 60 minutes protect from light at 25°C on shaker;
5. Remove liquid on magnetic frame, add 100μL prepared Detection Reagent B, and incubate 30 minutes protect from light at 25°C on shaker;
6. Wash plate once on magnetic frame;
7. Add 100μL sheath fluid, swirl for 10 minutes, read on the machine.

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Magazine Citations
Endocrine Journal Relationship between serum secreted frizzled-related protein 4 levels and the first-phase of glucose-stimulated insulin secretion in individuals with different glucose tolerance PubMed: 26041556
Horm Metab Res. Secreted Frizzled-Related Protein 4 (SFRP4) is Elevated in Patients with Diabetes Mellitus Pubmed:26882051
Obesity (Silver Spring).  Reduced expression of chemerin in visceral adipose tissue associates with hepatic steatosis in patients with obesity pubmed:27768254
Obesity Longitudinal Increases in Adiposity Contribute to Worsening Adipokine Profile over Time in Mexican Americans Pubmed:29427376
Novel marker for gestational diabetes patent:US20180059122A1
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
American Journal of Cancer Research SFRP4 is a prognostic marker and correlated with Treg cell infiltration in pancreatic ductal adenocarcinoma Pubmed: 30906634
Diabetologia Effects of long-term exercise on plasma adipokine levels and inflammation-related gene expression in subcutaneous adipose tissue in sedentary dysglycaemic … Pubmed: 31011777
PLoS One First trimester secreted Frizzled-Related Protein 4 and other adipokine serum concentrations in women developing gestational diabetes mellitus Pubmed: 33206702
Gastric Cancer SFRP4 drives invasion in gastric cancer and is an early predictor of recurrence 33277667
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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