FRZB ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Frizzled Related Protein (FRZB) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Frizzled Related Protein (FRZB) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

FRE; frezzled; FRITZ; frizzled homolog-related; frizzled-related protein 1; FRP-3; FRZB1; FRZB-1; FRZB-PEN; FZRB; hFIZ; OS1; secreted frizzled-related protein 3; sFRP-3; SFRP3; SRFP3.

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. SFRP3/FRZB appears to be involved in limb skeletogenesis. Antagonist of Wnt8 signaling. Regulates chondrocyte maturation and long bone development.

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 Frizzled Related Protein (FRZB) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Frizzled Related Protein (FRZB) . 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 Frizzled Related Protein (FRZB), 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 Frizzled Related Protein (FRZB) 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) 89-97 93 EDTA Plasma (n=5) 85-97 90 Heparin Plasma (n=5) 94-102 98 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 89-97 93 Serum (n=5) 89-97 93 EDTA Plasma (n=5) 85-97 90 EDTA Plasma (n=5) 85-97 90 Heparin Plasma (n=5) 94-102 98 Heparin Plasma (n=5) 94-102 98

Component Amount Anti-FRZB Microplate 96 Wells (12 x 8 Well Strips) FRZB Lyophilized Standard 2 x 2 ng 100X Biotinylated FRZB Detector Antibody 120 uL Avidin/HRP Conjugate 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 TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-FRZB Microplate 96 Wells (12 x 8 Well Strips) Anti-FRZB Microplate 96 Wells (12 x 8 Well Strips) FRZB Lyophilized Standard 2 x 2 ng FRZB Lyophilized Standard 2 x 2 ng 100X Biotinylated FRZB Detector Antibody 120 uL 100X Biotinylated FRZB Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 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 TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

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