ELISA Kit for Secreted Frizzled Related Protein 4 (SFRP4) Rattus norvegicus (Rat) Sandwich ELISA

FRP-4; FRPHE; Frizzled protein, human endometrium

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  • ELISA Kit for Secreted Frizzled Related Protein 4 (SFRP4) Packages (Simulation)
  • ELISA Kit for Secreted Frizzled Related Protein 4 (SFRP4) Packages (Simulation)
  • ELISA Kit for Secreted Frizzled Related Protein 4 (SFRP4) Results demonstration
  • SEF878Ra.jpg Typical Standard Curve
  • Certificate ISO9001: 2008, ISO13485: 2003 Registered

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.

ELISA Kit for Secreted Frizzled Related Protein 4 (SFRP4)

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 Secreted Frizzled Related Protein 4 (SFRP4). Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Secreted Frizzled Related Protein 4 (SFRP4). 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 Secreted Frizzled Related Protein 4 (SFRP4), 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 Secreted Frizzled Related Protein 4 (SFRP4) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

Citations

  • Relationship between serum secreted frizzled-related protein 4 levels and the first-phase of glucose-stimulated insulin secretion in individuals with different glucose tolerancePubMed: 26041556
  • Secreted Frizzled-Related Protein 4 (SFRP4) is Elevated in Patients with Diabetes MellitusPubmed:26882051
  • Reduced expression of chemerin in visceral adipose tissue associates with hepatic steatosis in patients with obesitypubmed:27768254
  • Longitudinal Increases in Adiposity Contribute to Worsening Adipokine Profile over Time in Mexican AmericansPubmed:29427376
  • Novel marker for gestational diabetespatent:US20180059122A1
  • A diet high in sugar-sweetened beverage and low in fruits and vegetables is associated with adiposity and a pro-inflammatory adipokine profilePubmed: 30375290
  • SFRP4 is a prognostic marker and correlated with Treg cell infiltration in pancreatic ductal adenocarcinomaPubmed: 30906634
  • Effects of long-term exercise on plasma adipokine levels and inflammation-related gene expression in subcutaneous adipose tissue in sedentary dysglycaemic …Pubmed: 31011777
  • First trimester secreted Frizzled-Related Protein 4 and other adipokine serum concentrations in women developing gestational diabetes mellitusPubmed: 33206702
  • SFRP4 drives invasion in gastric cancer and is an early predictor of recurrence33277667
  • 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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