NENF ELISA Kit (Human)

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

Cell growth-inhibiting protein 47; cell immortalization-related protein 2; CIR2; neudesin; neuron-derived neurotrophic factor; Protein GIG47; SCIRP10; SCIRP10-related protein; secreted protein of unknown function; Spinal cord injury related protein 10; SPUF.

Acts as a neurotrophic factor in postnatal mature neurons enhancing neuronal survival. Promotes cell proliferation and neurogenesis in undifferentiated neural pro-genitor cells at the embryonic stage and inhibits differentiation of astrocyte. Its neurotrophic activity is exerted via MAPK1/ERK2, MAPK3/ERK1 and AKT1/AKT pathways. Neurotrophic activity is enhanced by binding to heme. Acts also as an anorexigenic neurotrophic factor that contributes to energy balance (By similarity) . Plays a role in the human tumorigenesis (PubMed:22748190) .

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 Neudesin Neurotrophic Factor (NENF) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Neudesin Neurotrophic Factor (NENF) . 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 Neudesin Neurotrophic Factor (NENF), 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 Neudesin Neurotrophic Factor (NENF) 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) 98-105 101 EDTA plasma (n=5) 83-98 94 heparin plasma (n=5) 93-101 97 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 98-105 101 serum (n=5) 98-105 101 EDTA plasma (n=5) 83-98 94 EDTA plasma (n=5) 83-98 94 heparin plasma (n=5) 93-101 97 heparin plasma (n=5) 93-101 97

Component Amount Anti-NENF Microplate 96 Wells (12 x 8 Well Strips) NENF Lyophilized Standard 2 x 10 ng 100X Biotinylated NENF 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-NENF Microplate 96 Wells (12 x 8 Well Strips) Anti-NENF Microplate 96 Wells (12 x 8 Well Strips) NENF Lyophilized Standard 2 x 10 ng NENF Lyophilized Standard 2 x 10 ng 100X Biotinylated NENF Detector Antibody 120 uL 100X Biotinylated NENF 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 Neudesin Neurotrophic Factor (NENF) . No significant cross-reactivity or interference between Neudesin Neurotrophic Factor (NENF) and analogues was observed.