Product Features
Product Name Human beta-NGF EasyTest™ ELISA Kit Assay Type Solid Phase Sandwich ELISA Format 96-well strip plate Assay Process One wash step, two incubations Assay Length 1.5 hours Sample Volume Serum , Plasma, 50µL Detection Method Colorimetric Sensitivity < 1 pg/mL Assay Range 31.2-2000pg/mL Reactivity Human Cross-Reactivity No cross-reactivity observed with available related molecules Gene Names NGF; Beta-NGF; HSAN5B Gene ID 4803 Swissprot ID P01138
Recovery
Sample Type | Average % Recovery | Range % |
Serum | 95 | 89-107 |
Plasma | 103 | 99-107 |
Linearity
Sample Type | Dilution | % of Expected |
Serum | 1:2 | 91 |
1:4 | 83 | |
1:8 | 71 | |
Plasma | 1:2 | 94 |
1:4 | 85 | |
1:8 | 89 |
Standard Curve: Human beta-NGF EasyTest™ ELISA Kit
Kit Components
• Pre-Coated 96-well Strip Microplate • Wash Buffer • Stop Solution • Assay Diluent • Standards • Bioaim human beta-NGF mix • TMB One-Step Substrate
Protocol Outline
1. Prepare all reagents, samples and standards as instructed in the manual. 2. Add 50 µl of standard or sample to each well. 3. Add 50 µl of Bioaim mix. 4. Incubate 1 h at RT. 5. Wash and add 100 µl of TMB One-Step Substrate Reagent to each well. 6. Incubate 15 min at RT. 7. Add 50 µl of Stop Solution to each well. 8. Read at 450 nm/570 nm.
Background
Nerve growth factor (NGF) was initially isolated in the mouse submandibular gland over three decades ago as a 7S complex composed of three non- covalently linked subunits, alpha, beta, and gamma. It is now known that both the alpha and gamma subunits of NGF are members of the kallikrein family of serine proteases. As its name suggests, NGF is involved primarily in the growth, as well as the maintenance, proliferation, and survival of nerve cells (neurons). In fact, NGF is critical for the survival and maintenance of sympathetic and sensory neurons, as they undergo apoptosis in its absence.
NGF binds with at least two classes of receptors: the tropomyosine receptor kinase A (TrkA) and low-affinity NGF receptor (LNGFR/p75NTR). When NGF binds to the TrkA receptor, it drives the homodimerization of the receptor, which in turn causes the autophosphorylation of the tyrosine kinase segment. This leads to the activation of PI 3-kinase, ras, and PLC signaling pathways.
Storage
The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
Regulatory Status
Human beta-NGF EasyTest™ ELISA Kit is for Research Use Only.