CDC42 Binding Protein Kinase beta (DMPK-Like) (CDC42BPB) (Active) Protein

Details zu Produkt Nr. ABIN411974, Anbieter: Anmelden zum Anzeigen
Proteinname
  • CDC42BPB
  • wu:fb98c02
  • wu:fc48b05
  • wu:fc90d12
  • si:dkeyp-93h6.2
  • MRCKb
  • MRCKB
  • CDC42 binding protein kinase beta
  • CDC42 binding protein kinase beta (DMPK-like)
  • CDC42BPB
  • cdc42bpb
  • Cdc42bpb
Spezies
Human
8
Quelle
Baculovirus infected Insect Cells
4
2
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 MRCK beta.
Produktmerkmale Protein Source: Baculovirus (Sf9 insect cells)
Reinheit SDS-PAGE: ≥90 %
Hintergrund Myotonic Dystrophy Kinase-Related cdc42-binding kinase β (MRCKb) belongs to the DMPK subfamily (1). The myotonic dystrophy kinase-related kinases and myotonic dystrophy kinase-related Cdc42 binding kinase (MRCK) are effectors of RhoA and Cdc42, respectively, where they are involved in actin cytoskeletal reorganization and neurite outgrowth (2). Effects of the repeat expansion on the DMPK gene may be responsible for muscle and heart features of Myotonic Dystrophy.
Alternate Names: MRCK, CDC42 binding protein kinase beta (DMPK-like), DMPK-like beta, Myotonic dystrophy kinase-related CDC42-binding kinase beta
Molekulargewicht 73.0 kDa
Gen-ID 9578
NCBI Accession NM_006035
Applikationshinweise Optimal working dilution should be determined by the investigator.
Kommentare

Activity Specification: The specific activity of MRCKB was determined to be 120 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 MRCK beta 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 Kinase Assay Buffer I diluted at a 1:4 ratio (5X dilution) with 50ng/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 Buffer components: 20 mM MOPS, pH
7. 2, 12.0 mM beta-glycerol-phosphate, 20 mM MgC1 2, 0 mM EGTA, 0 mM EDTA. Add 0.20 mM DTT to Kinase Assay Buffer prior to use. Substrate S6K synthetic peptide substrate (KRRRLASLR) diluted in distilled H2O to a final concentration of 0 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 MRCK beta, 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 MRCK beta Component 2. 0 µL of 0 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.
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 CDC42 Binding Protein Kinase beta (DMPK-Like) (CDC42BPB) (Active) protein (ABIN411974) CDC42 Binding Protein Kinase beta (DMPK-Like) (CDC42BPB) (Active) protein
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