Rat TrkA ELISA Kit PicoKine®

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.

Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 115 123 127 129 124 2.72 2.2% Sample 2 863 769 752 806 798 16.75 2.1% Sample 3 2241 2249 2369 2477 2334 72.35 3.1% Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Lots Lot 1 (pg/ml) Lot 2 (pg/ml) Lot 3 (pg/ml) Lot 4 (pg/ml) Mean (pg/ml) Standard Deviation CV (%) Sample 1 115 123 127 129 124 2.72 2.2% Sample 1 115 123 127 129 124 2.72 2.2% Sample 2 863 769 752 806 798 16.75 2.1% Sample 2 863 769 752 806 798 16.75 2.1% Sample 3 2241 2249 2369 2477 2334 72.35 3.1% Sample 3 2241 2249 2369 2477 2334 72.35 3.1%

High affinity nerve growth factor receptor;2.7.10.1; Neurotrophic tyrosine kinase receptor type 1; Slow nerve growth factor receptor; p140-TrkA; Trk-A; Ntrk1; Trk, Trka

Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 115 863 2241 117 796 2366 Standard deviation 4.14 27.62 87.4 4.8 35.82 87.54 CV (%) 3.6% 3.2 3.9% 4.1% 4.5% 3.7% Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 Mean (pg/ml) 115 863 2241 117 796 2366 Standard deviation 4.14 27.62 87.4 4.8 35.82 87.54 CV (%) 3.6% 3.2 3.9% 4.1% 4.5% 3.7% Intra-Assay Precision Inter-Assay Precision Intra-Assay Precision Inter-Assay Precision Sample 1 2 3 1 2 3 Sample 1 2 3 1 2 3 n 16 16 16 24 24 24 n 16 16 16 24 24 24 Mean (pg/ml) 115 863 2241 117 796 2366 Mean (pg/ml) 115 863 2241 117 796 2366 Standard deviation 4.14 27.62 87.4 4.8 35.82 87.54 Standard deviation 4.14 27.62 87.4 4.8 35.82 87.54 CV (%) 3.6% 3.2 3.9% 4.1% 4.5% 3.7% CV (%) 3.6% 3.2 3.9% 4.1% 4.5% 3.7%

Receptor tyrosine kinase involved in the development and the maturation of the central and peripheral nervous systems through regulation of proliferation, differentiation and survival of sympathetic and nervous neurons. High affinity receptor for NGF which is its primary ligand, it can also bind and be activated by NTF3/neurotrophin-3. However, NTF3 only supports axonal extension through NTRK1 but has no effect on neuron survival. Upon dimeric NGF ligand-binding, undergoes homodimerization, autophosphorylation and activation. Recruits, phosphorylates and/or activates several downstream effectors including SHC1, FRS2, SH2B1, SH2B2 and PLCG1 that regulate distinct overlapping signaling cascades driving cell survival and differentiation. Through SHC1 and FRS2 activates a GRB2-Ras-MAPK cascade that regulates cell differentiation and survival. Through PLCG1 controls NF-Kappa-B activation and the transcription of genes involved in cell survival. Through SHC1 and SH2B1 controls a Ras- PI3 kinase-AKT1 signaling cascade that is also regulating survival. In absence of ligand and activation, may promote cell death, making the survival of neurons dependent on trophic factors. .

Description|Quantity Pre-coated 96-well strip microplate|1 Standard|2 vials Biotinylated antibody (100x) |100ul Avidin-Biotin-Peroxidase Complex (100x) |100ul Sample Diluent|30ml Antibody Diluent|12ml Avidin-Biotin-Peroxidase Diluent|12ml Color Developing Reagent (TMB) |10ml Stop Solution|10ml Wash Buffer (25x) |20ml Adhesive plate sealers|4

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cell membrane ; Single-pass type I membrane protein . Early endosome membrane; Single-pass type I membrane protein. Late endosome membrane; Single-pass type I membrane protein. Internalized to endosomes upon binding of NGF or NTF3 and further transported to the cell body via a retrograde axonal transport. Localized at cell membrane and early endosomes before nerve growth factor (NGF) stimulation. Recruited to late endosomes after NGF stimulation. Colocalized with RAPGEF2 at late endosomes.

Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.038 0.066 0.1 0.16 0.299 0.568 1.193 2.053 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.038 0.066 0.1 0.16 0.299 0.568 1.193 2.053 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 Concentration (pg/ml) 0 78 156 312 625 1250 2500 5000 O.D. 0.038 0.066 0.1 0.16 0.299 0.568 1.193 2.053 O.D. 0.038 0.066 0.1 0.16 0.299 0.568 1.193 2.053

Microplate Reader capable of reading absorbance at 450nm. Incubator. Automated plate washer (optional) . Pipettes and pipette tips capable of precisely dispensing 0.5 μl through 1 ml volumes of aqueous solutions. Multichannel pipettes are recommended for large amount of samples. Deionized or distilled water. 500ml graduated cylinders. Test tubes for dilution.