Mitogen-Activated Protein Kinase 11 (MAPK11) (Active) Protein

Details zu Produkt Nr. ABIN457518, Anbieter: Anmelden zum Anzeigen
Proteinname
  • ATMPK11
  • F22L4.10
  • F22L4_10
  • MAP kinase 11
  • zgc:86905
  • P38B
  • P38BETA2
  • PRKM11
  • SAPK2
  • SAPK2B
  • p38-2
  • p38Beta
  • P38b
  • Prkm11
  • Sapk2
  • Sapk2b
  • p38beta
  • p38beta2
  • p38b
  • sapk2
  • prkm11
  • sapk2b
  • mitogen-activated protein kinase 11
  • MAP kinase 11
  • mapk11
  • LOC9328118
  • MPK11
  • MAPK11
  • Mapk11
  • PTRG_02565
Spezies
Human
15
1
Quelle
Baculovirus infected Insect Cells
9
2
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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Verwendungszweck >90% Pure active Recombinant p38alpha.
Produktmerkmale Protein Source: Baculovirus (Sf9 insect cells)
Reinheit SDS-PAGE: ≥90 %
Hintergrund P38-β is a member of the p38 MAP kinase family and is activated by both proinflammatory cytokines and environmental stress (1). The p38-β is activated through its phosphorylation by MAP kinase kinases (MKKs), preferably by MKK6. Transcription factor ATF2/CREB2 has been shown to be a substrate of this kinase (2). Alternatively spliced transcript variants encoding the same protein have been observed.
Alternate Names: p38, mitogen-activated protein kinase 11
Molekulargewicht 71.0 kDa
Gen-ID 5600
NCBI Accession NM_002751
Pathways MAPK Signalweg, Neurotrophin Signalübertragung, Activation of Innate immune Response, Response to Water Deprivation, Regulation of Muscle Cell Differentiation, ER-Nucleus Signaling, Hepatitis C, Toll-Like Receptors Cascades, Signaling Events mediated by VEGFR1 and VEGFR2, Thromboxane A2 Receptor Signaling
Applikationshinweise Optimal working dilution should be determined by the investigator.
Kommentare

Activity Specification: The specific activity of P38BETA was determined to be 123 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 p38beta 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 p38beta, 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 p38beta. 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 bringing the final
6. volume up to 20 µL and incubate the mixture in a water bath at 30 °C for 15 minutes.
7. 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.
8. 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.
9. Count the radioactivity on the P81 paper in the presence of scintillation fluid in a scintillation counter. Step 10. 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, and 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 Mitogen-Activated Protein Kinase 11 (MAPK11) (Active) protein (ABIN457518) Mitogen-Activated Protein Kinase 11 (MAPK11) (Active) protein
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