GFER ELISA Kit (Mouse)

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

FAD-dependent sulfhydryl oxidase that regenerates the redox-active disulfide bonds in CHCHD4/MIA40, a chaperone essential for disulfide bond formation and protein folding in the mitochondrial intermembrane space. The reduced form of CHCHD4/MIA40 forms a transient intermolecular disulfide bridge with GFER/ERV1, resulting in regeneration of the essential disulfide bonds in CHCHD4/MIA40, while GFER/ERV1 becomes re-oxidized by donating electrons to cytochrome c or molecular oxygen (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 Growth Factor, Augmenter Of Liver Regeneration (GFER) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Growth Factor, Augmenter Of Liver Regeneration (GFER) . 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 Growth Factor, Augmenter Of Liver Regeneration (GFER), 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 Growth Factor, Augmenter Of Liver Regeneration (GFER) 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) 93-101 96 _x005F EDTA plasma (n=5) 78-101 84 _x005F heparin plasma (n=5) 97-105 101 _x005F Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 93-101 96 serum (n=5) 93-101 96 EDTA plasma (n=5) 78-101 84 EDTA plasma (n=5) 78-101 84 heparin plasma (n=5) 97-105 101 heparin plasma (n=5) 97-105 101

Component Amount _x005F Anti-Gfer Microplate 96 Wells (12 x 8 Well Strips) _x005F Gfer Lyophilized Standard 2 x 5 ng _x005F 100X Biotinylated Gfer 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-Gfer Microplate 96 Wells (12 x 8 Well Strips) Anti-Gfer Microplate 96 Wells (12 x 8 Well Strips) Gfer Lyophilized Standard 2 x 5 ng Gfer Lyophilized Standard 2 x 5 ng 100X Biotinylated Gfer Detector Antibody 120 uL 100X Biotinylated Gfer 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 Growth Factor, Augmenter Of Liver Regeneration (GFER) ._x005F No significant cross-reactivity or interference between Growth Factor, Augmenter Of Liver Regeneration (GFER) and analogues was observed.