Human Secreted Frizzled-Related Protein 1(SFRP1) ELISA Kit

Instructions
Code CSB-E15074h
Size 96T,5×96T,10×96T
Trial Size 24T ELISA kits trial application
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Product Details

Description

The Human Secreted Frizzled-Related Protein 1 (SFRP1) ELISA kit is a sandwich immunoassay specifically designed and validated for the quantitative detection of SFRP1 in multiple biological solutions, including serum, plasma, cell culture supernates, tissue homogenates, or cell lysates. It is not intended for diagnostic use. This assay kit was designed and optimized for cell biology research use in humans. The kit has undergone rigorous quality control in multiple parameters, including sensitivity, specificity, precision, linearity, recovery, and inter-batch difference. Refer to the product instructions for more details.

This assay employs the quantitative sandwich enzyme immunoassay technique, in which SFRP1 in the samples or standards are sandwiched between pre-coated SFRP1 antibody and Biotin-conjugated SFRP1 antibody. HRP-avidin is then added to the wells. Following a wash to remove any unbound reagent, the TMB substrate solution is added to the wells and color develops in proportion to the amount of SFRP1 bound in the initial step. The color development is stopped upon adding the stop solution, and the intensity of the color is measured at 450 nm via a microplate reader. The levels of SFRP1 in the samples can be determined by referring to the O.D. (optical density) of the samples to the standard curve.

SFRP1 is a negative regulator of Frizzled receptors and Wnt pathway activation. It is abundantly expressed in the heart. As a biphasic modulator of Wnt signaling, SFRP1 counteracts Wnt-induced effects at high concentrations and promotes them at lower concentrations. SFRP1 has been classified as a tumor suppressor due to the absence of its encoding gene in a variety of human cancers. This may result in carcinogenesis as a consequence of the dysregulation of cell proliferation, migration, and invasion.

Target Name secreted frizzled-related protein 1
Alternative Names Frizzled related protein 1 ELISA Kit; FRP 1 ELISA Kit; FRP ELISA Kit; FRP-1 ELISA Kit; FRP1 ELISA Kit; FrzA ELISA Kit; SARP 2 ELISA Kit; SARP-2 ELISA Kit; SARP2 ELISA Kit; Secreted apoptosis related protein 2 ELISA Kit; Secreted apoptosis-related protein 2 ELISA Kit; Secreted frizzled related protein 1 ELISA Kit; Secreted frizzled related protein ELISA Kit; Secreted frizzled-related protein 1 ELISA Kit; SFRP 1 ELISA Kit; sFRP-1 ELISA Kit; SFRP1 ELISA Kit; SFRP1_HUMAN ELISA Kit
Abbreviation SFRP1
Uniprot No. Q8N474
Species Homo sapiens (Human)
Sample Types serum, plasma, cell culture supernates, tissue homogenates, cell lysates
Detection Range 3.12 pg/mL-200 pg/mL
Sensitivity 0.78 pg/mL
Assay Time 1-5h
Sample Volume 50-100ul
Detection Wavelength 450 nm
Research Area Cell Biology
Assay Principle quantitative
Measurement Sandwich
Precision
Intra-assay Precision (Precision within an assay): CV%<8%
Three samples of known concentration were tested twenty times on one plate to assess.
Inter-assay Precision (Precision between assays): CV%<10%
Three samples of known concentration were tested in twenty assays to assess.
Linearity
To assess the linearity of the assay, samples were spiked with high concentrations of human SFRP1 in various matrices and diluted with the Sample Diluent to produce samples with values within the dynamic range of the assay.
 SampleSerum(n=4)
1:100Average %83
Range %80-86
1:200Average %95
Range %90-101
1:400Average %96
Range %93-100
1:800Average %90
Range %85-96
Recovery
The recovery of human SFRP1 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay as directed in the Sample Preparation section.
Sample TypeAverage % RecoveryRange
Serum (n=5) 9690-102
EDTA plasma (n=4)8582-90
Typical Data
These standard curves are provided for demonstration only. A standard curve should be generated for each set of samples assayed.
pg/mlOD1OD2AverageCorrected
2002.217 2.269 2.243 2.060
1001.758 1.787 1.773 1.590
501.331 1.357 1.344 1.161
250.932 0.968 0.950 0.767
12.50.613 0.637 0.625 0.442
6.250.445 0.469 0.457 0.274
3.120.303 0.328 0.316 0.133
00.182 0.184 0.183  
Materials provided
  • A micro ELISA plate --- The 96-well plate has been pre-coated with an anti-human SFRP1 antibody. This dismountable microplate can be divided into 12 x 8 strip plates.
  • Two vials lyophilized standard ---Dilute a bottle of the standard at dilution series, read the OD values, and then draw a standard curve.
  • One vial Biotin-labeled SFRP1 antibody (100 x concentrate) (120 μl/bottle) ---Act as the detection antibody.
  • One vial HRP-avidin (100 x concentrate) (120 μl/bottle) ---Bind to the detection antibody and react with the TMB substrate to make the solution chromogenic.
  • One vial Biotin-antibody Diluent (15 ml/bottle) ---Dilute the Biotin-antibody.
  • One vial HRP-avidin Diluent (15 ml/bottle) ---Dilute the HRP-avidin solution.
  • One vial Sample Diluent (50 ml/bottle)---Dilute the sample to an appropriate concentration.
  • One vial Wash Buffer (25 x concentrate) (20 ml/bottle) ---Wash away unbound or free substances.
  • One vial TMB Substrate (10 ml/bottle) ---Act as the chromogenic agent. TMB interacts with HRP, eliciting the solution turns blue.
  • One vial Stop Solution (10 ml/bottle) ---Stop the color reaction. The solution color immediately turns from blue to yellow.
  • Four Adhesive Strips (For 96 wells) --- Cover the microplate when incubation.
  • An instruction manual
Materials not provided
  • A microplate reader capable of measuring absorbance at 450 nm, with the correction wavelength set at 540 nm or 570 nm.
  • An incubator can provide stable incubation conditions up to 37°C±5°C.
  • Centrifuge
  • Vortex
  • Squirt bottle, manifold dispenser, or automated microplate washer
  • Absorbent paper for blotting the microtiter plate
  • 50-300ul multi-channel micropipette
  • Pipette tips
  • Single-channel micropipette with different ranges
  • 100ml and 500ml graduated cylinders
  • Deionized or distilled water
  • Timer
  • Test tubes for dilution
Troubleshooting
and FAQs
ELISA kit FAQs
Storage Store at 2-8°C. Please refer to protocol.
Lead Time 3-5 working days

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Target Background

Function
(From Uniprot)
Soluble frizzled-related proteins (sFRPS) function as modulators of Wnt signaling through direct interaction with Wnts. They have a role in regulating cell growth and differentiation in specific cell types. SFRP1 decreases intracellular beta-catenin levels (By similarity). Has antiproliferative effects on vascular cells, in vitro and in vivo, and can induce, in vivo, an angiogenic response. In vascular cell cycle, delays the G1 phase and entry into the S phase (By similarity). In kidney development, inhibits tubule formation and bud growth in metanephroi (By similarity). Inhibits WNT1/WNT4-mediated TCF-dependent transcription.
Gene References into Functions
  1. active involvement in placental development and important role in pathology of pregnancy PMID: 28738713
  2. SFRP1 gene was epigenetically silenced in glioblastomas when compared to low astrocytoma grades, which may suggest that the lack of its protein is involved in astrocytoma progression. PMID: 30394013
  3. It has been found that miR-27a-3p modulated the Wnt/beta-catenin signaling pathway to promote epithelial-mesenchymal transition in oral squamous carcinoma stem cells by down-regulating SFRP1. PMID: 28425477
  4. sFRP1 overexpression in gastric cancer cells leads to increased cell proliferation and angiogenesis. PMID: 28169332
  5. Study determined that SFRP1 is downregulated during tumor formation probably by DNA methylation, and associates consistently with tumor suppressive functions. PMID: 28218291
  6. co-transfection of sFRP1 siRNA with miR- 191 inhibitor sequence, it was found that the Wnt/beta-catenin signaling pathway was reactivated and the colony formation ability of RBE cells was restored. PMID: 30235453
  7. WNT5b rs3803164, and SFRP1 rs3242 were significantly associated with lumbar spine BMD. PMID: 30055306
  8. lost SFRP1 expression-induced Wnt/beta-catenin signaling due to the hypermethylation of the SFRP1 promoter could associate with keloid development PMID: 29455276
  9. miR-1301-3p promoted the expansion of prostate cancer stem cells by inhibiting GSK3beta and SFRP1, and activating the Wnt pathway. PMID: 29358129
  10. Methylation level of the SFRP1 promoter region is increased in patients with RCC compared to normal controls and might be involved in the occurrence and development of RCC. PMID: 29731501
  11. preliminary data show loss of SFRP1 protein expression caused by the SFRP1 promoter hypermethylation in a subset of high-grade serous ovarian carcinomas PMID: 29146058
  12. Loss of SFRP1 likely contributes to tumor progression by altering the expression of a critical transcription factor in both the epithelium and the immune system. PMID: 28687085
  13. Authors found that downregulation of HP1alpha can decrease H3K9me3 enrichment and DNA methylation rate of secreted frizzled-related protein 1 (SFRP1) promoter, resulting in restoring the expression of SFRP1. PMID: 27385214
  14. KL and SFRP1 methylation were more predominant in nasopharyngeal tumors. PMID: 28812223
  15. Results indicate that SFRP1 rs7832767 C > T, CTNNB1 rs2293303 C > T, and WISP1 rs16893344 C > T were all strongly correlated with myocardial infarction (MI) susceptibility. PMID: 27391264
  16. miR-1254 promotes lung cancer cell proliferation by targeting the expression of SFRP1. PMID: 28605875
  17. The expression of Dicer negatively correlated with that of SFRP1 and it appeared to promote CCA cell proliferation. PMID: 28230864
  18. H2O2-induced SFRP1 gene demethylation contributes to H2O2-induced apoptosis in human U251 glioma cells. PMID: 28398872
  19. Data suggest that microRNA miR-27a may activate the Wnt/beta-catenin signaling pathway by negatively regulating SFRP1 to promote the proliferation, migration and invasion of breast cancer (BC) cells. PMID: 28099945
  20. Data indicate that microRNA miR-744 activated Wnt/beta-catenin pathway by targeting multiple negative regulators of Wnt/beta-catenin signaling, including SFRP1, GSK3beta, TLE3 and NKD1, and that NKD1 is a major functional target of miR-744. PMID: 28107193
  21. Our results indicate that DKK1 and SFRP1 may be potentially useful biomarkers for evaluating the beneficial effects of long-term exercise on physical fitness and metabolism as well as the prognosis of patients with cancer. PMID: 28178355
  22. MiR-1-3p suppresses the proliferation, invasion and migration of bladder cancer cells by up-regulating SFRP1 expression. PMID: 28268231
  23. SFRP1 mRNA expression was reduced by both resistant starch and polydextrose. Resistant starch and polydextrose did not affect SFRP1 methylation or alter the expression of 10 microRNAs predicted to target SFRP1. PMID: 28077379
  24. Taken together, our results suggest that canonical Wnt signaling and its antagonist, sFRP1, regulate proliferation of human CSCs. Furthermore, excess sFRP1 in elderly patients causes CSC aging. PMID: 28435069
  25. Genetic variants in the 3' untranslated region of sFRP1 gene is associated with risk of gastric cancer. PMID: 27739564
  26. This study suggested mechanistic relationship between miR-940 and Wnt/beta-catenin in the development and progression of pancreatic carcinoma through regulation of GSK3beta and sFRP1. PMID: 27459115
  27. In prostate cancer decreased SFRP1 expression can be due to occupancy of H3K27me3 or to promoter DNA hypermethylation. PMID: 27570179
  28. SFRP1 expression was significantly lower in cutaneous squamous cell carcinoma tissues than that in tissues of control subjects. Histopathological differentiation in CSCC negatively correlated with SFRP1 expression. PMID: 27420949
  29. miR-1303 promotes neuroblastoma (NB) proliferation by targeting GSK3beta and SFRP1, and might be a target for NB therapy. PMID: 27434867
  30. The secreted frizzled-related protein and disheveled protein family members appear to be actively involved in the pathogenesis of primary testicular germ cell tumors. PMID: 27599467
  31. Results show that SNPs rs4736958 and rs7832767 of SFRP1 gene were significantly associated with tuberculosis susceptibility and might influence the expression levels of inflammatory markers of tuberculosis patients in a Chinese Han population. PMID: 26643984
  32. Down-regulation of SFRP1 is associated with recurrence in Endometrioid Adenocarcinoma. PMID: 26397159
  33. SFRP1 downregulation may result from epigenetic silencing and is associated with worse survival in nasopharyngeal carcinoma PMID: 26276746
  34. in gliomas there was correlation of SFRP1 methylation with tumor grade and patients' survival PMID: 26337424
  35. Hepatocellular carcinoma patients with high SFRP1 expression in tumor tissue had a significantly better overall survival rate. PMID: 26851021
  36. Reduced SFRP1 expression is associated with acute myeloid leukemia. PMID: 25347432
  37. Hypermethylation of the selected markers (MAL, PRIMA1, PTGDR and SFRP1) can result in reduced gene expression and may contribute to the formation of colorectal cancer. PMID: 26482433
  38. SFRP1 is a direct target of MiR-27a in the regulation of glioma invasion and cell migration. PMID: 26164457
  39. In human mammary epithelial cells with either SFRP1 knocked down (TERT-siSFRP1) and rescued SFRP1 expression (MCF7-SFRP1), estrogen signaling is augmented. PMID: 25809273
  40. The increased placental expression of sFRP1 seen in smokers impairs fetal growth by inhibiting WNT signaling and trophoblast proliferation. PMID: 26413870
  41. miR-27a induced gene silencing contributes to bone metabolism in hFOB cells in vitro, which is partly affected by the post-transcriptional regulation of sFRP1, during osteoblast proliferation, apoptosis and differentiation. PMID: 24625818
  42. While SFRP1 re-expression generally mediated distinct patterns of transcriptionally induced or repressed genes in BT20 and SKBR3 cells, BDNF was a SFRP1-induced gene in both cell lines. PMID: 25036590
  43. sFRP1 protein displays antineoplastic activity. PMID: 25432334
  44. Knockdown of SFRP1 expression in RT112 cells resulted only in marginal effects. PMID: 25732201
  45. the DNA methylation levels of OPCML and SFRP1 could be potential biomarkers for diagnosis of CCA with high specificity, sensitivity, and accuracy, in particular for biopsy specimens PMID: 25652468
  46. Sfrp1 contributes to coordination of visual axon growth in retinal ganglion cells. PMID: 25788689
  47. study indicates that increased expression of both SFRP1 and SFRP3 may contribute to the pathogenesis of IUGR placental dysfunction, whereas the loss of these proteins may be involved in the development of human trophoblastic tumors. PMID: 25046226
  48. Polymorphisms in several genes involved in the Wnt signaling pathway were associated with hepatic fibrosis or inflammation risk in HCV-infected males. PMID: 24386373
  49. Coexpression of SFRP1 and WIF1, rather than SFRP1 or WIF1 alone, could be used, together with low TNM stage, as a prognostic predictor of favorable outcomes in CRC. PMID: 24949429
  50. Our study provided the first evidence that SNPs in CTNNB1 and SFRP1 were associated with tuberculosis risk, and genetic variants in Wnt signaling pathway might participate in genetic susceptibility to tuberculosis in Chinese Han population. PMID: 24695522

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Subcellular Location Secreted
Protein Families Secreted frizzled-related protein (sFRP) family
Tissue Specificity Widely expressed. Absent from lung, liver and peripheral blood leukocytes. Highest levels in heart and fetal kidney. Also expressed in testis, ovary, fetal brain and lung, leiomyomal cells, myometrial cells and vascular smooth muscle cells. Expressed in f
Database Links

HGNC: 10776

OMIM: 604156

KEGG: hsa:6422

STRING: 9606.ENSP00000220772

UniGene: Hs.213424

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