|Application ||WB, LCI|
|Reactivity||Human, Mouse, Rat|
|Calculated MW||87868 Da|
|Homology||Mouse - identical; human - 13/14 amino acid residues identical.|
|Other Names||High affinity nerve growth factor receptor, Neurotrophic tyrosine kinase receptor type 1, Slow nerve growth factor receptor, p140-TrkA, Trk-A, Ntrk1, Trk, Trka|
|Related products for control experiments||Control peptide antigen (supplied with the antibody free of charge).|
|Target/Specificity||Peptide (C)ETMRHG(S)LRLNQPTH, corresponding to amino acid residues 342-356 of rat TrkA (Accession P35739). Extracellular, 2nd immunoglobulin-like domain.|
|Peptide Confirmation||Confirmed by amino acid analysis.|
|Format||Affinity purified antibody, lyophilized powder|
|Reconstitution||50 µl or 0.2 ml deionized water, depending on the sample size.|
|Antibody Concentration After Reconstitution||0.8 mg/ml.|
|Buffer After Reconstitution||Phosphate buffered saline (PBS), pH 7.4, 1% BSA, 0.025% NaN3.|
|Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Storage After Reconstitution||The reconstituted solution can be stored at 4ºC for up to 2 weeks. For longer periods, small aliquots should be stored at -20ºC or below. Avoid multiple freezing and thawing. The further dilutions should be made using a carrier protein such as BSA (1%). Centrifuge all antibody preparations before use (10000 × g 5 min).|
|Control Antigen Storage Before Reconstitution||Lyophilized powder can be stored intact at room temperature for several weeks. For longer periods, it should be stored at -20°C.|
|Control Antigen Reconstitution||100 µl water.|
|Control Antigen Storage After Reconstitution||-20ºC.|
|Preadsorption Control||1 µg peptide per 1 µg antibody.|
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Provided below are standard protocols that you may find useful for product applications.
NGF was the first neurotrophin to be discovered, more than 50 years ago. Three other proteins have been added to the list: BDNF, NT-3 and NT-4 (NT-4/5). These neurotrophins bind two groups of receptors. The p75NTR receptor is common to all four neurotrophins and is a member of the tumor necrosis factor receptor family. The tropomyosin-related kinase (TrK) receptors are receptor tyrosine kinases (RTKs) and three receptors form this family: TrkA, TrkB, and TrkC1. As mentioned above, the p75NTR receptor binds to all neurotrophins with similar affinities while the Trk receptors are the ones to display the selectivity for the neurotrophins. TrkA is activated by NGF binding, TrkB by that of BDNF and NT-4, while TrkC is stimulated by the binding of NT-3 1. All three Trk receptors are highly expressed in the mammalian brain in very distinct regions and are also expressed in the peripheral nervous system2-4. Cholinergic neurons in the basal forebrain exclusively express TrkA. All three Trk receptors are expressed in the hippocampus. Motor and sensory neurons in the peripheral nervous system express Trk receptors. Interestingly, while Trk receptors are not essential for development, knockout mice die shortly after birth. Indeed, TrkA-deficient mice demonstrate a significant decrease in cholinergic neurons and sympathetic neurons in the trigeminal, superior cervical and dorsal root ganglia5-7. Trk receptors are type I membrane proteins (i.e. span the membrane with one transmembrane region|) and have many motifs in the extracellular region, including cell-adhesion domains, three tandem leucine rich motifs flanked by two clusters of cysteines. In the membrane proximal region of the receptor there are also two immunoglobulin-like domains8. The intracellular region of the protein contains the kinase domain, responsible for activating downstream effectors as well as many adaptor site regions important for the binding/interaction of signaling molecules. The binding of neurotrophins to Trk receptors promotes receptor dimerization resulting in kinase activation. Activated Trk receptors then phosphorylate a cascade of signaling molecules including the Ras/ERK, PI3K/Akt pathways and PLC-γ1. Activated Trk receptors also create internal docking sites for other signaling adaptor proteins to bind to8. Splice variants of TrkA, TrkB and TrkC have been observed. These splice isoforms are mainly affected in the tyrosine kinase domain of the receptor lying in the cytoplasm. Endocytosis is an important signaling trait of Trk receptors. Following neurotrophin binding to the Trk receptor, the receptor complex is then internalized via endocytosis in order to terminate signaling. However, in the axonal compartment of neurons the internalization process of the neurotrophin complexed to the receptor is part of the signaling process and is important for activating transcription processes in the nucleus9,10. Abgent is pleased to offer a highly specific antibody directed against an epitope located at the extracellular domain of rat TrkA receptor. Anti-TrkA (extracellular) antibody (#AG1416) can be used in western blot, immunohistochemical, immunocytochemical and indirect flow cytometry applications, and has been designed to recognize TrkA from mouse, rat and human samples.
References 1. Webster, N.J.G. and Pirrung, M.C. (2008) BMC Neurosci. 9, S1. 2. Quartu, M. et al. (2003) Int. J. Dev. Neurosci. 21, 309. 3. Aoki, C. et al. (2000) J. Neurosci. Res. 59, 454. 4. Yan, Q. et al. (1997) J. Comp. Neurol. 378, 135. 5. Crowley, C. et al. (1994) Cell 76, 1001. 6. Smeyne, R.J. et al. (1994) Nature 368, 246. 7. Fagan, A.M. et al. (1997) J. Neurosci. 17, 7644. 8. Patapoutian, A. and Reichardt, L.F. (2001) Curr. Opin. Neurobiol. 11, 272. 9. Schecterson, L.C. and Bothwell, M. (2010) Develop. Neurobiol. 70, 332. 10. Grimes, M.L. et al. (1996) J. Neurosci. 16, 7950.
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