Fyn-Related Kinase (FRK) (Active) Protein

Details zu Produkt Nr. ABIN411970, Anbieter: Anmelden zum Anzeigen
Proteinname
  • BK2
  • NGK1
  • FRK
  • GTK
  • PTK5
  • RAK
  • BSK
  • BSK/IYK
  • C85044
  • potassium voltage-gated channel subfamily A member 2
  • Fyn-related kinase
  • fyn related Src family tyrosine kinase
  • tyrosine-protein kinase FRK
  • fyn-related kinase
  • fyn-related Src family tyrosine kinase
  • Kcna2
  • FRK
  • frk
  • LOC100540680
  • Frk
Spezies
Human
10
1
Quelle
Baculovirus infected Insect Cells
6
2
1
1
1
Protein-Typ
Recombinant
Biologische Aktivität
Active
Applikation
Functional Studies (Func), SDS-PAGE (SDS), Western Blotting (WB)
Optionen
Hersteller
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Hersteller Produkt- Nr.
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Verwendungszweck >90% Pure active Recombinant FRK.
Produktmerkmale Protein Source: Baculovirus (Sf9 insect cells)
Reinheit SDS-PAGE: ≥90 %
Hintergrund FRK (fyn-related kinase) or Rak is a nuclear tyrosine kinase and member of the Src sub-family. Restricted expression of FRK is detected in a broad range of cell lines with highest levels in epithelial cells. Increased expression of FRK has been shown in breast and renal cell carcinoma cell lines. In addition the retinoblastoma tumor susceptibility gene product pRb associates with FRK in vitro and in vivo (1). Overexpression of FRK in β-cells from the pancreas increases the susceptibility of these cells to β-cell-toxic events (hallmark of Type I diabetes)(2).
Alternate Names: FRK, fyn-related kinase, FYN-related kinase, Nuclear tyrosine protein kinase RAK, Protein-tyrosine kinase 5
Molekulargewicht 60.0 kDa
Gen-ID 2444
NCBI Accession NM_002031
Forschungsgebiet Kinases/Phosphatases
Applikationshinweise Optimal working dilution should be determined by the investigator.
Kommentare

Activity Specification: The specific activity of FRK was determined to be 997 nmol /min/mg as per activity assay protocol.

Protokoll Note: these are suggested working dilutions and it is recommended that the researcher perform a serial dilution of Active FRK 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, 12.0 mM beta-glycerol-phosphate, 20 mM MgC1 2, 20 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 FRK, 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 FRK. Component 2. 10 µL of 1 mg/mL stock solution of substrate
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.
Beschränkungen Nur für Forschungszwecke einsetzbar
Format Liquid
Konzentration 0.1 mg/mL
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.
Konservierungsmittel Dithiothreitol (DTT)
Vorsichtsmaßnahmen This product contains dithiothreitol (DTT): a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handhabung Centrifuge the vial prior to opening.
Lagerung -80 °C
Informationen zur Lagerung -80°C
Haltbarkeit 12 months
Bilder des Herstellers
Western Blotting (WB) image for Fyn-Related Kinase (FRK) (Active) protein (ABIN411970) Fyn-Related Kinase (FRK) (Active) protein
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