NGF High Sensitivity ELISA Kit (Human)

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

Beta-nerve growth factor; Beta-NGF; HSAN5; nerve growth factor (beta polypeptide) ; nerve growth factor, beta subunit; NGFB; pro-nerve growth factor long.

Nerve growth factor is important for the development and maintenance of the sympathetic and sensory nervous systems. Extracellular ligand for the NTRK1 and NGFR receptors, activates cellular signaling cascades through those receptor tyrosine kinase to regulate neuronal proliferation, differentiation and survival. Inhibits metalloproteinase dependent proteolysis of platelet glycoprotein VI (PubMed:20164177) .

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 High Sensitive Nerve Growth Factor (NGF) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to High Sensitive Nerve Growth Factor (NGF) . 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 High Sensitive Nerve Growth Factor (NGF), 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 High Sensitive Nerve Growth Factor (NGF) 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) 96-104 101 EDTA Plasma (n=5) 98-105 102 Heparin Plasma (n=5) 99-105 102 Matrix Recovery range (%) Average (%) Matrix Recovery range (%) Average (%) Serum (n=5) 96-104 101 Serum (n=5) 96-104 101 EDTA Plasma (n=5) 98-105 102 EDTA Plasma (n=5) 98-105 102 Heparin Plasma (n=5) 99-105 102 Heparin Plasma (n=5) 99-105 102

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