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EK000004-EK0308: Rat BDNF ELISA Kit
Range 31.2pg/ml-2000pg/ml
Sensitivity < 2pg/ml
Specificity: no detectable cross-reactivity with any other cytokine.
Application: for quantitive detection of rat BDNF in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: four months at 4°C and eight months at -20°C.
Background
Brain-derived neurotrophic factor (BDNF) is a prosurvival factor induced by cortical neurons that is necessary for survival of striatal neurons in the brain. It is a secreted protein with the molecular weight of 27.8kDa, consisting of 247 amino acids. It is known to promote neuronal survival and differentiation. BDNF shares substantial amino acid sequence identity with nerve growth factor (NGF). BDNF and neurotrophin-3 (NT-3) are two recently cloned neurotrophic factors that are homologous to NGF. mRNA products of the BDNF and NT-3 genes are detected in the adult human brain, suggesting that these proteins are involved in the maintenance of the adult nervous system.1 BDNF and other neurotrophins are critically involved in long-term potentiation (LTP). BDNF-mediated LTP is induced postsynaptically.2 BDNF has trophic effects on serotonergic (5-HT) neurons in the central nervous system.3 BDNF has an essential maintenance function in the regulation of anxiety-related behavior and in food intake through central mediators in both the basal and fasted state.4 It plays a role in treating breathing disorders such as respiratory insufficiency after spinal injury.5 The mature form of BDNF is identical in all mammals examined, and the gene encoding human BDNF to chromosome 11, band p13. 6
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. Jones, K. R.; Reichardt, L. F. Molecular cloning of a human gene that is a member of the nerve growth factor family. Proc. Nat. Acad. Sci. 87: 8060-8064, 1990.
2. Kovalchuk, Y.; Hanse, E.; Kafitz, K. W.; Konnerth, A. Postsynaptic induction of BDNF-mediated long-term potentiation. Science 295: 1729-1734, 2002.
3. Lyons, W. E.; Mamounas, L. A.; Ricaurte, G. A; Coppola, V.; Reid, S. W.; Bora, S. H.; Wihler, C.; Koliatsos, V. E.; Tessarollo, L. Brain-derived neurotrophic factor-deficient mice develop aggressiveness and hyperphagia in conjunction with brain serotonergic abnormalities. Proc. Nat. Acad. Sci. 96: 15239-15244, 1999.
4. Rios, M.; Fan, G.; Fekete, C.; Kelly, J.; Bates, B.; Kuehn, R.; Lechan, R. M.; Jaenisch, R. Conditional deletion of brain-derived neurotrophic factor in the postnatal brain leads to obesity and hyperactivity. Molec. Endocr. 15: 1748-1757, 2001.
5. Baker-Herman, T. L.; Fuller, D. D.; Bavis, R. W.; Zabka, A. G.; Golder, F. J.; Doperalski, N. J.; Johnson, R. A.; Watters, J. J.; Mitchell, G. S. BDNF is necessary and sufficient for spinal respiratory plasticity following intermittent hypoxia. Nature Neurosci. 7: 48-55, 2004.
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