1. CELLULAR KINASE MTO pTrkA (Tyr490) and (Tyr674/675)

2. 2 Contents  General introduction  Signaling pathway  Assay format  Product specifications  Assay performances  Ordering information  Bibliography

3. 3 pTrkA (Tyr490) and (Tyr674/675): general introduction  TrkA is a neurotrophic tyrosine receptor kinase family that also includes TrkB and TrkC and is a high affinity receptor for NGF (nerve growth factor)  Specific neuronal cell survival, mitosis and differentiation are dependent on the activation of TrkA  The activation of TrkA by NGF is followed by the activation of growth and survival pathways by cytoplasmic proteins: SHC, PLCgamma1 and P13 kinase http://drugdiscoveryopinion.com  Specifically, phosphorylation at Tyr490 is required for Shc association and activation of the Ras-MAP kinase cascade. The phosphorylation at Tyr674/675 indicate TrkA kinase activity  Diseases in which the activation of TrkA is indicated are cancers of the breast, ovary, prostate, and thyroid  Recent animal studies involving activation of TrkA in relation to stimulated bladder activity suggest the utilization of TrkA as a biomarker  Alternate names: C80751, DKFZp781I14186, MTC, NTRK1, p140 Trk, p140-TrkA, Tkr, TRK, TRK1, TRKA, TRKA longest isoform, TRKA NGFR

4. 4 Associated therapeutic areas :  Oncology  Growth Factor Receptors  Neuroscience  Neutrophins Signal-transduction pathway of the TrkA tyrosine kinase receptor Other TrkA related pathways:  RAS signaling pathway  PI3K / Akt signaling pathway  MAPK signaling pathway

5. 5 p-TrkA (Tyr490) and (Tyr 674/675): assay format  Cell stimulation: triggers activation  Cell lysis: releases the phosphorylated kinase  Kinase detection: detects the phosphorylated TrkA in a sandwich assay format using an anti-phospho TrkA antibody labeled with Eu3+-Cryptate and an anti-Total TrkA antibody labeled with d2

6. 6 p-TrkA (Tyr490) and (Tyr 674/675): product specifications  Determination of phospho TrkA in cell lysates only when phosphorylated at Tyrosine 490 or Tyrosine 674/675  Assay robustness  Incubation 2 hrs at 37ºC and then transferred to ON at 4ºC  High assay window  Cross reactivity tested: human

7. 7 p-TRKA (Tyr 490) and (Tyr 674/675): assay performance

8. 8 pTRKA (Tyr490) and (Tyr674/675):ordering information Part numberDescription name Number of tests per kit List price 63ADK029PEGPhospho TRKA (Tyr490)500$1,450 63ADK029PEHPhospho TRKA (Tyr490)10,000$7,900 Part numberDescription name Number of tests per kit List price 63ADK030PEGPhospho TRKA (Tyr674/675)500$1,450 63ADK030PEHPhospho TRKA (Tyr674/675)10,000$7,900

9. 9 References:  The activated nerve growth factor receptor p-TrkA is selectively expressed in advanced- stage ovarian carcinoma Ødegaard E, Staff AC, Abeler VM, Kopolovic J, Onsrud M, Lazarovici P, Davidson B Department of Gynecology, Ulleval University Hospital, University of Oslo, N-0407 Oslo, Norway. Hum Pathol. 2007 Jan;38(1):140-6. Epub 2006 Sep 25Ødegaard EStaff ACAbeler VMKopolovic JOnsrud M Lazarovici PDavidson BHum Pathol.  Biomarkers in an Overactive Bladder, Alka A. Bhide, Rufus Cartwright, Vik Khullar, G. Alessandro Digesu International Urogynecology Journal, January 2013 Alka A. BhideRufus CartwrightVik KhullarG. Alessandro Digesu  The Crystal Structures of TrkA and TrkB Suggest Key Regions for Achieving Selective Inhibition, T. Bertrand 1 M. Kothe 3,J. Liu 3, A. Dupuy 1, A. Rak 1,P.F. Berne 2, S. Davis 3,T. Gladysheva 3,C Valtre 2, J.Y. Crenne 2, M. Mathieu Journal of Molecular Biology, Vol 423, Issue 3, 26 October 2012, Pages 439–453T. Bertrand 1 M. Kothe 3J. Liu 3A. Dupuy 1A. Rak 1P.F. Berne 2S. Davis 3T. Gladysheva 3C Valtre 2J.Y. Crenne 2M. Mathieu

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