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4638
TrkA and TrkB Antibody Sampler Kit
Primary Antibodies
Antibody Sampler Kit

TrkA and TrkB Antibody Sampler Kit #4638

Citations (2)
Simple Western™ analysis of tissue extract (0.1 mg/mL) from mouse brain using Trk (pan) (A7H6R) Rabbit mAb #92991. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from NIH/3T3, NIH/3T3-TrkA, and NIH/3T3-TrkB using TrkA (12G8) Rabbit mAb (upper) TrkB (80E3) Rabbit mAb Antibody #4603 (middle) and PLCγ1 (D9H10) XP® Rabbit mAb #5690 (lower).
Western blot analysis of extracts from neonatal mouse brain, rat brain and NIH/3T3/TrkB cells using TrkB (80E3) Rabbit mAb.
Western blot analysis of extracts from NIH/3T3 cells stably transfected with either TrkA or TrkB, left untreated or treated with NGF or BDNF, respectively, using Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9) Rabbit mAb (upper) and pooled TrkA/TrkB antibodies (lower).
Western blot analysis of extracts from NIH/3T3 cells stably transfected with TrkA or TrkB, and treated with NGF or BDNF, respectively, using Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb (upper) and pooled TrkA/TrkB Antibodies (lower).
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
Western blot analysis of extracts from mouse neonatal and rat fetal brain using Trk (pan) (A7H6R) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded NIH/3T3-TrkA (left), NIH/3T3-TrkB (middle) and NIH/3T3-TrkC (right) cell pellets using TrkA (12G8) Rabbit mAb.
Simple Western™ analysis of lysates (1.0 mg/mL) from Moue Brain tissue using TrkB (80E3) Rabbit mAb #4603. The virtual lane view (left) shows the target band (as indicated) at 1:10 and 1:50 dilutions of primary antibody. The corresponding electropherogram view (right) plots chemiluminescence by molecular weight along the capillary at 1:10 (blue line) and 1:50 (green line) dilutions of primary antibody. This experiment was performed under reducing conditions on the Jess™ Simple Western instrument from ProteinSimple, a BioTechne brand, using the 12-230 kDa separation module.
Western blot analysis of extracts from untreated or NGF-treated 3T3/TrkA and PC12 cells using Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9) Rabbit mAb.
Western blot analysis of extracts from untreated or NGF-treated PC12 cells using Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb.
Western blot analysis of extracts from M0-91, KM12 cells and mouse neonatal brain using Trk (pan) (A7H6R) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using TrkA (12G8) Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).
Immunoprecipitation of mouse neonatal brain extracts. Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Trk (pan) (A7H6R) Rabbit mAb. Western blot analysis was performed using Trk (pan) (A7H6R) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human prostate carcinoma using TrkA (12G8) Rabbit mAb.
To Purchase # 4638
Cat. # Size Qty. Price
4638T
1 Kit  (5 x 20 microliters)

Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9) Rabbit mAb 4619 20 µl
  • WB
  • IP
H R 140 Rabbit IgG
Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb 4621 20 µl
  • WB
  • IP
H R 140 Rabbit IgG
TrkA (12G8) Rabbit mAb 2510 20 µl
  • WB
  • IHC
H 140 Rabbit IgG
TrkB (80E3) Rabbit mAb 4603 20 µl
  • WB
H M R 90, 140 Rabbit IgG
Trk (pan) (A7H6R) Rabbit mAb 92991 20 µl
  • WB
  • IP
H M R 120-140 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Rab Goat 

Product Description

The TrkA and TrkB Antibody Sampler Kit provides an economical means to investigate the Trk family of tyrosine kinase receptors. The kit contains enough primary and secondary antibodies to perform four Western blots with each antibody.

Specificity / Sensitivity

Both total TrkA and TrkB antibodies detect endogenous levels of their respective Trk receptors and do not cross-react with related proteins. The Trk (pan) (A7H6R) Rabbit mAb detects endogenous levels of total TrkA, TrkB, and TrkC proteins. However, the antibody may preferentially detect TrkA over TrkB and TrkB over TrkC. Phospho-TrkA (Tyr490)/TrkB (Tyr516) (C35G9) Rabbit mAb detects endogenous levels of TrkA and TrkB only when phosphorylated at the indicated sites; this antibody may cross-react with Bcr-Abl phosphorylated at an unknown tyrosine residue. Phospho-TrkA (Tyr674/675)/TrkB (Tyr706/707) (C50F3) Rabbit mAb detects endogenous levels of TrkA and TrkB only when phosphorylated at the indicated sites; this antibody may cross-react with a protein of ~150 kDa phosphorylated at an unknown tyrosine residue.

Source / Purification

Total monoclonal antibodies are produced by immunizing animals with synthetic peptides (KLH-coupled) surrounding Arg220 of human TrkA, Pro50 of human TrkB, and Tyr791 of human TrkA. Activation state monoclonal antibodies are produced by immunizing animals with synthetic phosphopeptides (KLH-coupled) corresponding to residues surrounding Tyr490 of human TrkA or Tyr674/674 of human TrkA.

Background

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).

  1. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
  2. Segal, R.A. and Greenberg, M.E. (1996) Annu Rev Neurosci 19, 463-89.
  3. Stephens, R.M. et al. (1994) Neuron 12, 691-705.
  4. Marsh, H.N. et al. (2003) J Cell Biol 163, 999-1010.
  5. Obermeier, A. et al. (1993) EMBO J 12, 933-41.
  6. Obermeier, A. et al. (1994) EMBO J 13, 1585-90.
  7. Arevalo, J.C. et al. (2001) Oncogene 20, 1229-34.
  8. Reuther, G.W. et al. (2000) Mol Cell Biol 20, 8655-66.
  9. Greco, A. et al. (1997) Genes Chromosomes Cancer 19, 112-23.
  10. Pierotti, M.A. and Greco, A. (2006) Cancer Lett 232, 90-8.
  11. Lagadec, C. et al. (2009) Oncogene 28, 1960-70.
  12. Greco, A. et al. (2010) Mol Cell Endocrinol 321, 44-9.
  13. Ødegaard, E. et al. (2007) Hum Pathol 38, 140-6.
  14. Huang, E.J. and Reichardt, L.F. (2003) Annu Rev Biochem 72, 609-42.
  15. Geiger, T.R. and Peeper, D.S. (2005) Cancer Res 65, 7033-6.
  16. Han, L. et al. (2007) Med Hypotheses 68, 407-9.
  17. Aoyama, M. et al. (2001) Cancer Lett 164, 51-60.
  18. Desmet, C.J. and Peeper, D.S. (2006) Cell Mol Life Sci 63, 755-9.

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