Mitogen-Activated Protein Kinase 8 (MAPK8) (Active) Protein

Details for Product No. ABIN593493, Supplier: Log in to see
Protein Name
  • BSK
  • BSK/DJNK
  • Bsk
  • CG5680
  • D-JNK
  • D-junk
  • DBSK/JNK
  • DJNK
  • DJNK/bsk
  • Dmel\\CG5680
  • JNK
  • JNK/SAPK
  • Jnk
  • Junk
  • SAPKa
  • c-Jun
  • dJNK
  • jnk
  • JNK-46
  • JNK1
  • JNK1A2
  • JNK21B1/2
  • PRKM8
  • SAPK1
  • SAPK1c
  • AI849689
  • Prkm8
  • jnk1
  • sapk1
  • T10F20.15
  • mapk8
  • zgc:112379
  • basket
  • mitogen-activated protein kinase 8
  • mitogen-activated protein kinase 8b
  • Protein JNK-1
  • bsk
  • MAPK8
  • Mapk8
  • mapk8
  • ATMPK8
  • mapk8b
  • jnk-1
Origin
Human
17
7
3
1
1
1
Source
Baculovirus infected Insect Cells
11
5
4
4
3
2
1
Protein Type
Recombinant
Biological Activity
Active
Application
Functional Studies (Func), SDS-PAGE (SDS), Western Blotting (WB)
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Purpose >90% Pure active Recombinant JNK1.
Characteristics Protein Source: Baculovirus (Sf9 insect cells)
Purity SDS-PAGE: ≥90 %
Background JNK1 is a member of the MAP kinase group that is activated by dual phosphorylation at thr and tyr residues during exposure to stress such as UV irradiation. JNK1 binds to the c-Jun transactivation domain and phosphorylates it on Ser-63 and Ser-73 (1). JNK1 has been shown to play an important role in disease processes. Activation of JNK1 results in defects in myotube viability and integrity leading to dystrophic myofiber destruction (2). JNK1 activity is also abnormally elevated in obesity and removal of JNK1 results in decreased adiposity and significantly improved insulin sensitivity.
Alternate Names: JNK, mitogen-activated protein kinase 8, MAP kinase p49 3F12, Stress-activated protein kinase 1b, Stress-activated protein kinase JNK3, c-Jun N-terminal kinase 3
Molecular Weight 71.0 kDa
Gene ID 5599
UniProt P45983
Research Area Immunology, Innate Immunity
Pathways MAPK Signaling, WNT Signaling, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Hepatitis C, Toll-Like Receptors Cascades, Signaling of Hepatocyte Growth Factor Receptor
Application Notes Optimal working dilution should be determined by the investigator.
Comment

Activity Specification: The specific activity of JNK1 was determined to be 124 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 JNK1 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 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 I, pH 7.2 Buffer components: 20 mM MOPS, 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 ATF2 substrate prepared in buffer (50 mM Tris-HCl, pH
7. 2, 50 mM NaC1 2, 0 mM EDTA and 0.20 mM DTT) to a final concentration of 0.0 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 JNK1, 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 JNK1. Component 2. 10 µL of 0.0 mg/mL ATF2 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 bring 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 mg/mL
Buffer Recombinant proteins in storage buffer (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 Mitogen-Activated Protein Kinase 8 (MAPK8) (Active) protein (ABIN593493) Mitogen-Activated Protein Kinase 8 (MAPK8) (Active) protein
Product cited in: Sury, McShane, Hernandez-Miranda, Birchmeier, Selbach et al.: "Quantitative proteomics reveals dynamic interaction of c-Jun N-terminal kinase (JNK) with RNA transport granule proteins splicing factor proline- and glutamine-rich (Sfpq) and non-POU ..." in: Molecular & cellular proteomics : MCP, Vol. 14, Issue 1, pp. 50-65, 2015 (PubMed).

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