Product Pathways - PathScan ELISA
PathScan® Phospho-TrkB (panTyr) Sandwich ELISA Kit #7108
|7108S||1 Kit (96 assays)||---||In Stock||---|
|7108||carrier free and custom formulation / quantity||email request|
When ordering five or more kits, please contact us for processing time and pricing at firstname.lastname@example.org.
Important Ordering Details: Product is assembled upon order to ensure maximum activity. United States: Please allow up to two weeks for your order to be processed and shipped. Outside of the United States: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.
Already purchased this product? Write a Review.
|Kit Includes||Volume||Solution Color|
|TrkB Ab Coated Microwells||96 tests|
|Biotinylated P-Tyrosine Detection Ab||11 ml||Green|
|HRP-linked Streptavidin||11 ml||Red|
|TMB Substrate #7004||11 ml||Colorless|
|STOP Solution #7002||11 ml||Colorless|
|Sealing Tape||2 sheets|
|ELISA Wash Buffer (20X)||25 ml||Colorless|
|ELISA Sample Diluent||25 ml||Blue|
|Cell Lysis Buffer (10X) #9803||15 ml||Yellowish|
Note: 12 8-well modules – Each module is designed to break apart for 8 tests.
Storage: Kit should be stored at 4°C with the exception of Cell Lysis Buffer, which is stored at –20°C (packaged separately).
The PathScan® Phospho-TrkB (panTyr) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects transfected levels of TrkB when phosphorylated on tyrosine residues. A TrkB mouse antibody has been coated onto the microwells. After incubation with cell lysates, TrkB (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a biotinylated phospho-tyrosine detection antibody is added to detect tyrosine phosphorylation of the captured TrkB protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of TrkB phosphorylated on tyrosines.
Antibodies in kit are custom formulations specific to kit.
Specificity / Sensitivity
CST's PathScan® Phospho-TrkB (panTyr) Sandwich ELISA Kit #7108 detects transfected levels of phospho-TrkB protein when tyrosines are phosphorylated(see Figure 1). The kit sensitivity is shown in figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
ELISA - Western correlation
Figure 1: Treatment of 3T3/TrkB cells with BDNF (#3897) stimulates tyrosine-phosphorylation of TrkB, detected by PathScan® Phospho-TrkB (panTyr) Sandwich ELISA Kit #7108, but does not affect the level of total TrkB detected by PathScan® Total TrkB Sandwich ELISA Kit #7106. Absorbance at 450 nm is shown in the top figure, while the corresponding Western blots using Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb #4621 (right panel) or TrkB (80E3) Rabbit mAb #4603 (left panel), are shown in the bottom figure. The human TrkB is transfected and expressed in NIH/3T3 cells.
The family of Trk receptor tyrosine kinases consists of TrkA, TrkB, and TrkC. While the sequence of these family members is highly conserved, they are activated by different neurotrophins: TrkA by NGF, TrkB by BDNF or NT4, and TrkC by NT3 (1). Neurotrophin signaling through these receptors regulates a number of physiological processes, such as cell survival, proliferation, neural development, and axon and dendrite growth and patterning (1). In the adult nervous system, the Trk receptors regulate synaptic strength and plasticity. TrkA regulates proliferation and is important for development and maturation of the nervous system (2). Phosphorylation at Tyr490 is required for Shc association and activation of the Ras-MAP kinase cascade (3,4). Residues Tyr674/675 lie within the catalytic domain, and phosphorylation at these sites reflects TrkA kinase activity (3-6). Point mutations, deletions, and chromosomal rearrangements (chimeras) cause ligand-independent receptor dimerization and activation of TrkA (7-10). TrkA is activated in many malignancies including breast, ovarian, prostate, and thyroid carcinomas (8-13). Research studies suggest that expression of TrkA in neuroblastomas may be a good prognostic marker as TrkA signals growth arrest and differentiation of cells originating from the neural crest (10).
The phosphorylation sites are conserved between TrkA and TrkB: Tyr490 of TrkA corresponds to Tyr512 in TrkB, and Tyr674/675 of TrkA to Tyr706/707 in TrkB of the human sequence (14). TrkB is overexpressed in tumors, such as neuroblastoma, prostate adenocarcinoma, and pancreatic ductal adenocarcinoma (15). Research studies have shown that in neuroblastomas, overexpression of TrkB correlates with an unfavorable disease outcome when autocrine loops signaling tumor survival are potentiated by additional overexpression of brain-derived neurotrophic factor (BDNF) (16-18). An alternatively spliced truncated TrkB isoform lacking the kinase domain is overexpressed in Wilms’ tumors and this isoform may act as a dominant-negative regulator of TrkB signaling (17).
- Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
- Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89.
- Stephens, R.M. et al. (1994) Neuron 12, 691-705.
- Marsh, H.N. et al. (2003) J Cell Biol 163, 999-1010.
- Obermeier, A. et al. (1993) EMBO J 12, 933-41.
- Obermeier, A. et al. (1994) EMBO J 13, 1585-90.
- Arevalo, J.C. et al. (2001) Oncogene 20, 1229-34.
- Reuther, G.W. et al. (2000) Mol Cell Biol 20, 8655-66.
- Greco, A. et al. (1997) Genes Chromosomes Cancer 19, 112-23.
- Pierotti, M.A. and Greco, A. (2006) Cancer Lett 232, 90-8.
- Lagadec, C. et al. (2009) Oncogene 28, 1960-70.
- Greco, A. et al. (2010) Mol Cell Endocrinol 321, 44-9.
- Ødegaard, E. et al. (2007) Hum Pathol 38, 140-6.
- Huang, E.J. and Reichardt, L.F. (2003) Annu. Rev. Biochem. 72, 609-642.
- Geiger, T.R. and Peeper, D.S. (2005) Cancer Res 65, 7033-6.
- Han, L. et al. (2007) Med Hypotheses 68, 407-9.
- Aoyama, M. et al. (2001) Cancer Lett 164, 51-60.
- Desmet, C.J. and Peeper, D.S. (2006) Cell Mol Life Sci 63, 755-9.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 7111 PathScan® Phospho-TrkB (Tyr516) Sandwich ELISA Kit
- 7118 PathScan® Phospho-TrkB (Tyr706/707) Sandwich ELISA Kit
- 7106 PathScan® Total TrkB Sandwich ELISA Kit
- 4619 Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9) Rabbit mAb
- 4621 Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb
- 4603 TrkB (80E3) Rabbit mAb
- 4607 TrkB (80G2) Rabbit mAb
- 4606 TrkB Antibody
- 4609 Trk (pan) (C17F1) Rabbit mAb
- 3897 Human Brain-Derived Neurotrophic Factor (BDNF)
- 9803 Cell Lysis Buffer (10X)
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.