Neurotrophic Tyrosine Kinase, Receptor, Type 2 (NTRK2) (Active) Protein

Details for Product No. ABIN411965, Supplier: Log in to see
Protein Name
  • trkb
  • trkb-b
  • xtrkb
  • si:ch211-220b11.1
  • trkB2
  • NTRK2
  • ntrk2
  • gp145-trkb
  • TRKB
  • GP145-TrkB
  • trk-B
  • AI848316
  • C030027L06Rik
  • Tkrb
  • trkB
  • RATTRKB1
  • TRKB1
  • Trk-B
  • TrkB
  • neurotrophic receptor tyrosine kinase 2
  • neurotrophic receptor tyrosine kinase 2 L homeolog
  • neurotrophic tyrosine kinase, receptor, type 2b
  • neurotrophic receptor tyrosine kinase 2 S homeolog
  • neurotrophic tyrosine kinase, receptor, type 2
  • NTRK2
  • ntrk2.L
  • ntrk2b
  • ntrk2.S
  • ntrk2
  • Ntrk2
Origin
Human
28
5
2
2
1
Source
Baculovirus infected Insect Cells
11
11
7
2
2
1
1
1
1
1
Protein Type
Recombinant
Biological Activity
Active
Application
Functional Studies (Func), SDS-PAGE (SDS), Western Blotting (WB)
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Purpose >80% Pure active Recombinant TrkB.
Characteristics Protein Source: Baculovirus (Sf9 insect cells)
Purity SDS-PAGE: ≥80 %
Background TRKB is a member of the neurotrophic tyrosine receptor kinase (NTRK) family. TRKB is the high affinity catalytic receptor for several "neurotrophins", which are small protein growth factors that induce the survival and differentiation of distinct cell populations (1). TRKB is a membrane-bound receptor that, upon neurotrophin binding, phosphorylates itself and members of the MAPK pathway (2). Signalling through TRKB leads to cell differentiation. Mutations in the TRKB gene have been associated with obesity and mood disorders.
Alternate Names: TrkB, neurotrophic tyrosine kinase, receptor, type 2, GP145-TrkB, Neurotrophic tyrosine kinase receptor type 2, TrkB tyrosine kinase, Tropomyosin-related kinase B
Molecular Weight 67.0 kDa
Gene ID 4915
NCBI Accession NM_001018064
Research Area Cancer
Pathways RTK Signaling, Neurotrophin Signaling Pathway, cAMP Metabolic Process, Skeletal Muscle Fiber Development, Feeding Behaviour, Dicarboxylic Acid Transport
Application Notes Optimal working dilution should be determined by the investigator.
Comment

Activity Specification: The specific activity of TRKB was determined to be 74 nmol /min/mg as per activity assay protocol.

Protocol Note: these are suggested working dilutions and it is recommended that the researcher perform a serial dilution of Active SYK for optimal results). [ 32 P]-ATP Assay Cocktail Prepare 250μM [ 32 P]-ATP Assay Cocktail in a designated radioactive working area by adding the following components: 150 µL of 10 mM ATP Stock Solution, 100 µL [ 32P ]- ATP (0 mCi/100 µL), 5.70 ml of Kinase Assay Buffer). Store 0 ml aliquots at -20 °C. Kinase Dilution Buffer, pH 7.2 Kinase Assay Buffer II diluted at a 1:4 ratio (5X dilution) with 50 ng/µL BSA solution. 10 mM ATP Stock Solution Prepare ATP stock solution by dissolving 50 mg of ATP in 10 ml of Kinase Assay Buffer. Store 200 µL aliquots at -20 °C. Kinase Assay Buffer II, pH 7.2 Buffer components: 20 mM MOPS pH 7.2, 12.0 mM beta-glycerol-phosphate, 20 mM MgC1 2, 12.0 mM MnC1 2, 0 mM EGTA, 0 mM EDTA. Add 0.20 mM DTT to Kinase Assay Buffer prior to use. Substrate Poly (Glu:Tyr, 4:1) synthetic peptide substrate diluted in distilled H2O to a final concentration of 1 mg/mL. Assay Protocol Step 1. Thaw [ 32 P]-ATP Assay Cocktail in shielded container in a designated radioactive working area. Step 2. Thaw the Active TrkB, Kinase Assay Buffer, Substrate and Enzyme Dilution Buffer on ice. Step 3. In a pre-cooled microfuge tube, add the following reaction components bringing the initial reaction volume up to 20 µL: Component 1. 10 µL of diluted Active TrkB. Component 2. 0 µL of 1 mg/mL stock solution of substrate. Component 3. 0 µL distilled H2O (4 °C)
4. Set up the blank control as outlined in step 3, excluding the addition of the substrate. Replace the substrate with an equal volume of distilled H2O.
5. Initiate the reaction by the addition of 0 µL [ 32 P]-ATP Assay Cocktail bringing the final volume up to 20 µL and incubate the mixture in a water bath at 30 °C for 15 minutes.
6. After the 15 minute incubation period, terminate the reaction by spotting 20 µL of the reaction mixture onto individual pre-cut strips of phosphocellulose P81 paper.
7. Air dry the pre-cut P81 strip and sequentially wash in a 1 % phosphoric acid solution (dilute 10 ml of phosphoric acid and make a 1L solution with distilled H2O) with constant gentle stirring. It is recommended that the strips be washed a total of 3 intervals for approximately 10 minutes each.
8. Count the radioactivity on the P81 paper in the presence of scintillation fluid in a scintillation counter.
9. Determine the corrected cpm by removing the blank control value (see Step 4) for each sample and calculate the kinase specific activity as outlined below.
Restrictions For Research Use only
Format Liquid
Concentration 0.1 μg/μL
Buffer Recombinant protein stored in 50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.25 mM DTT, 0.1 mM EGTA, 0.1 mM EDTA, 0.1 mM PMSF, 25 % glycerol.
Preservative Dithiothreitol (DTT)
Precaution of Use This product contains dithiothreitol (DTT): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Centrifuge the vial prior to opening.
Storage -80 °C
Storage Comment -80°C
Expiry Date 12 months
Supplier Images
Western Blotting (WB) image for Neurotrophic Tyrosine Kinase, Receptor, Type 2 (NTRK2) (Active) protein (ABIN411965) Neurotrophic Tyrosine Kinase, Receptor, Type 2 (NTRK2) (Active) protein
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