ELISA Kit for Secreted Frizzled Related Protein 4 (SFRP4)

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).
No significant cross-reactivity or interference between Secreted Frizzled Related Protein 4 (SFRP4) and analogues was observed.

Recovery

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

Matrix Recovery range (%) Average(%)
serum(n=5) 84-102 98
EDTA plasma(n=5) 84-104 97
heparin plasma(n=5) 96-103 101

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Secreted Frizzled Related Protein 4 (SFRP4) 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) 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) 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) 83-93% 80-101% 78-101% 98-105%
EDTA plasma(n=5) 84-96% 78-101% 86-101% 89-104%
heparin plasma(n=5) 97-104% 80-103% 83-93% 93-101%

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
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
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