CFH ELISA Kit (Human)

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

Adrenomedullin binding protein; age-related maculopathy susceptibility 1; AHUS1; AMBP1; ARMD4; ARMS1; beta-1H; beta-1-H-globulin; CFHL3; complement factor H; factor H; factor H-like 1; FH; FHL1; H factor 1; H factor 1 (complement) ; H factor 2 (complement) ; HF; HF1; HF2; HUS.

Factor H functions as a cofactor in the inactivation of C3b by factor I and also increases the rate of dissociation of the C3bBb complex (C3 convertase) and the (C3b) NBB complex (C5 convertase) in the alternative complement pathway.

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 Complement Factor H (CFH) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Complement Factor H (CFH) . 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 Complement Factor H (CFH), 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 Complement Factor H (CFH) 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 (%) _x005F Serum (n=5) 92-105 98 _x005F EDTA Plasma (n=5) 87-94 91 _x005F Heparin Plasma (n=5) 83-91 87 _x005F Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 92-105 98 Serum (n=5) 92-105 98 EDTA Plasma (n=5) 87-94 91 EDTA Plasma (n=5) 87-94 91 Heparin Plasma (n=5) 83-91 87 Heparin Plasma (n=5) 83-91 87

Component Amount _x005F Anti-CFH Microplate 96 Wells (12 x 8 Well Strips) _x005F CFH Lyophilized Standard 2 x 300 ng _x005F 100X Biotinylated CFH Detector Antibody 120 uL _x005F Avidin/HRP Conjugate 120 uL _x005F Standard Diluent 1 x 20 mL _x005F Detector Antibody Diluent 1 x 12 mL _x005F Conjugate Diluent 1 x 12 mL _x005F 30X Wash Buffer 1 x 20 mL _x005F TMB Substrate 1 x 9 mL _x005F Stop Solution 1 x 6 mL _x005F Component Amount Component Amount Anti-CFH Microplate 96 Wells (12 x 8 Well Strips) Anti-CFH Microplate 96 Wells (12 x 8 Well Strips) CFH Lyophilized Standard 2 x 300 ng CFH Lyophilized Standard 2 x 300 ng 100X Biotinylated CFH Detector Antibody 120 uL 100X Biotinylated CFH 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 Complement Factor H (CFH) ._x005F No significant cross-reactivity or interference between Complement Factor H (CFH) and analogues was observed.