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EK000033-EK0471: Rat NGF ELISA Kit
Range 15.6pg/ml-1000pg/ml
Sensitivity < 1 pg/ml
Specificity: no detectable cross-reactivity with BDNF, GDNF, CNTF, NT3, and NT4.
Application: for quantitative detection of rat NGF in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Nerve growth factor (NGF) is a polypeptide involved in the regulation of growth and differentiation of sympathetic and certain sensory neurons. NGF is thought to have a profound effect on the development and maintenance of sympathetic and embryonic sensory neurones. NGF activity isolated from the male mouse submaxillary gland (MSG) consists of three types of subunits, alpha, beta and gamma, which specifically interact to form a 7S, approximately 130,000-molecular weight (Mr) complex. The 7S complex contains two identical 118-amino acid beta-chains, which are solely responsible for the nerve growth-stimulating activity of NGF.1 NGF, which is expressed by inflammatory cells and effects changes that lead to increased neural responsiveness, could be a pivotal mediator in allergic rhinitis.2 The standard product used in this kit is mouse 2.5S NGF, which is a dimmer linking with two polypeptide chains of 120 amino acids.
Principle
The ELISA Kit is based on standard sandwich enzyme-linked immune-sorbent assay technology. The target specific antibodies are precoated onto 96-well plates. The target from the sample is bound to the microwell. The biotinylated target specific detection antibodies are added to the microwells and followed by washing with the PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex is added and unbound conjugates are washed away with the PBS or TBS buffer. TMB, the HRP (horseradish peroxidase) substrate, is used to visualize color change resulting from the HRP enzymatic reaction. TMB is catalyzed by HRP to produce the blue color. The color changes into yellow after the acidic stop solution is added. The density of the yellow color is proportional to the target amount from the sample captured in the microwells.
Reference
1. Ullrich, A.; Gray, A.; Berman, C.; Dull, T. J. Human beta-nerve growth factor gene sequence highly homologous to that of mouse. Nature 303: 821-825, 1983.
2. Sanico, A. M.; Stanisz, A. M.; Gleeson, T. D.; Bora, S.; Proud, D.; Bienenstock, J.; Koliatsos, V. E.; Togias, A. Nerve growth factor expression and release in allergic inflammatory disease of the upper airways. Am. J. Resp. Crit. Care Med. 161: 1631-1635, 2000.
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