SFRP4 ELISA Kit (Rat)

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%

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 (By similarity) . SFRP4 plays a role in bone morphogenesis (By similarity) . May also act as a regulator of adult uterine morphology and function (PubMed:12063187) . May also increase apoptosis during ovulation possibly through modulation of FZ1/FZ4/WNT4 signaling (PubMed:12960062) . Has phosphaturic effects by specifically inhibiting sodium-dependent phosphate uptake (By similarity) .

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.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Matrix Recovery range (%) Average (%) Serum (n=5) 84-102 98 EDTA Plasma (n=5) 84-104 97 Heparin Plasma (n=5) 96-103 101 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 84-102 98 Serum (n=5) 84-102 98 EDTA Plasma (n=5) 84-104 97 EDTA Plasma (n=5) 84-104 97 Heparin Plasma (n=5) 96-103 101 Heparin Plasma (n=5) 96-103 101

Component Amount Anti-SFRP4 Microplate 96 Wells (12 x 8 Well strips) SFRP4 Lyophilized Standard 2 100X Biotinylated SFRP4 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL Component Amount Component Amount Anti-SFRP4 Microplate 96 Wells (12 x 8 Well strips) Anti-SFRP4 Microplate 96 Wells (12 x 8 Well strips) SFRP4 Lyophilized Standard 2 SFRP4 Lyophilized Standard 2 100X Biotinylated SFRP4 Detector Antibody 1 x 120 uL 100X Biotinylated SFRP4 Detector Antibody 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL 100X Avidin-HRP Conjugate 1 x 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL

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.