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EK000137-EK0847: Rat TrkA ELISA Kit
Range 156pg/ml-10000pg/ml
Sensitivity < 10pg/ml
Specificity: No detectable cross-reactivity with any other trk.
Application: For quantitative detection of rat TRKA in sera, plasma, body fluids, tissue lysates or cell culture supernates.
Expiration: Four months at 4°C and eight months at -20°C.
Background
Tropomyosin receptor kinase A (TrkA) are efficacious in attenuating skeletal pain.1 TrkA mutants were able to activate signaling cascades and were even more efficient in promoting neurite outgrowth than the wild-type receptor.2 TrkA is part of a sub-family of protein kinases which includes TrkB and TrkC. Also, there are other neurotrophic factors structurally related to NGF: BDNF (for Brain-Derived Neurotrophic Factor), NT-3 (for Neurotrophin-3) and NT-4 (for Neurotrophin-4). While TrkA mediates the effects of NGF, TrkB is bound and activated by BDNF, NT-4, and NT-3. Further, TrkC binds and is activated by NT-3.3 TrkA receptor was found in keratoconus-affected corneas, along with an increased level of repressor isoform of Sp3 transcription factor.4 The standard product used in this kit is extracellular part (A33-P418) of recombinant rat TRKA, consisting of 386 amino acids with the molecular weight of 69KDa.
Principle
TRKA ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. Rat TRKA specific-specific polyclonal antibodies were precoated onto 96-well plates. The rat specific detection polyclonal antibodies were biotinylated. The test samples and biotinylated detection antibodies were added to the wells subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the rat TRKA amount of sample captured in plate.
Reference
1. Casta?eda-Corral G, Jimenez-Andrade JM, Bloom AP, Taylor RN, Mantyh WG, Kaczmarska MJ, Ghilardi JR, Mantyh PW. Neuroscience. 2011 Mar 31;178:196-207. Epub 2011 Jan 26.
2. Georgieva MV, de Pablo Y, Sanchis D, Comella JX, Llovera M. J Neurochem. 2011 Feb 19.
3. Benito-Guti轵rez E, Garcia-Fern囝dez J, Comella JX (February 2006). "Origin and evolution of the Trk family of neurotrophic receptors". Mol. Cell. Neurosci. 31 (2): 179?2.
4. Lambiase A, Merlo D, Mollinari C, Bonini P, Rinaldi AM, D' Amato M, Micera A, Coassin M, Rama P, Bonini S, Garaci E (November 2005). "Molecular basis for keratoconus: lack of TrkA expression and its transcriptional repression by Sp3". Proc. Natl. Acad. Sci. U.S.A. 102 (46): 16795?00.
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