ELISA Kit for Secreted Frizzled Related Protein 1 (SFRP1)

FRP1; FRP; FRP-1; FrzA; SARP2; Secreted apoptosis-related protein 2

Specificity

This assay has high sensitivity and excellent specificity for detection of Secreted Frizzled Related Protein 1 (SFRP1).
No significant cross-reactivity or interference between Secreted Frizzled Related Protein 1 (SFRP1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Secreted Frizzled Related Protein 1 (SFRP1) and the recovery rates were calculated by comparing the measured value to the expected amount of Secreted Frizzled Related Protein 1 (SFRP1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 80-97 83
EDTA plasma(n=5) 92-101 96
heparin plasma(n=5) 79-99 95

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Secreted Frizzled Related Protein 1 (SFRP1) 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 1 (SFRP1) 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 1 (SFRP1) 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) 92-101% 86-96% 91-99% 91-98%
EDTA plasma(n=5) 80-92% 85-99% 79-105% 86-101%
heparin plasma(n=5) 96-103% 79-97% 82-96% 94-103%

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
BMC Musculoskeletal Disorders Biomarkers and cytokines of bone turnover: extensive evaluation in a cohort of patients with ankylosing spondylitis PubMed: 23025387
Exp Cell Res Wnt Inhibition is Dysregulated in Gliomas and its Re-Establishment Inhibits Proliferation and Tumor Sphere Formation PubMed: 26712519
BMC Musculoskeletal Disorders Correlation of blood bone turnover biomarkers and Wnt signaling antagonists with AS, DISH,OPLL, and OYL. pubmed:28153008
PloS one Effects of exercise training on circulating levels of Dickkpof-1 and secreted frizzled-related protein-1 in breast cancer survivors: A pilot single-blind randomized controlled trial. pubmed:28178355
PLoS One Effects of exercise training on circulating levels of Dickkpof-1 and secreted frizzled-related protein-1 in breast cancer survivors: A pilot single-blind randomized controlled trial 10.1371
Journal of Experimental & Clinical Cancer Research PHF21B overexpression promotes cancer stem cell-like traits in prostate cancer cells by activating the Wnt/β-catenin signaling pathway pubmed:28645312
JBMR Plus Low serum levels of DKK2 predict incident low‐impact fracture in older women Doi: 10.1002/jbm4.10179
Nature Communications Immunological history governs human stem cell memory CD4 heterogeneity via the Wnt signaling pathway Pubmed: 32041953
acta pharmacologica sinica sFRP1 protects H9c2 cardiac myoblasts from doxorubicin-induced apoptosis by inhibiting the Wnt/PCP-JNK pathway Pubmed: 32238888
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