Attention: We have moved. Merck Millipore products are no longer available for purchase on MerckMillipore.com.Learn More

171539 AMPK (α2/β1/γ1), His•Tag®, Human, Recombinant,S. frugiperda

171539
Purchase on Sigma-Aldrich

Descripción

Replacement Information

Products

Número de referenciaEmbalaje Cant./Env.
171539-5UG Ampolla de plást. 5 μg
Description
OverviewFull length, recombinant, human AMPK consisting of subunits α2/β1/γ1, each fused at the C-terminus to a His•Tag® sequence and co-expressed in S. frugiperda insect cells using a baculovirus expression system. AMPK is involved in sensing energy levels in the cell that result from changes in AMP levels. AMPK is an upstream regulator of mTOR activation. The α-subunit contains a kinase domain, the β-subunit contains a glycogen binding domain, and the γ-subunit contains the regulatory AMP binding domain. The active complex is useful in the study of kinase activity regulation and inhibitor screening.
Catalogue Number171539
Brand Family Calbiochem®
SynonymsAMP Kinase (α2/β1/γ1)
Application Data
The activity of AMPK (α2/β1/γ1) was measured as outlined in the Recommended Reaction Conditions.
References
ReferencesForetz, M., et al. 2005. Diabetes 54, 1331.
Viollet, B., et al. 2003. Biochem. Soc. Trans. 31, 216.
Product Information
Unit of DefinitionKinase activity was measured as the molar amount of phosphate incorporated into SAMS peptide substrate (HMRSAMSGLHLVKRR) at 30°C using a final concentration of 50 µM ATP.
FormLiquid
FormulationIn 150 mM NaCl, 50 mM Tris-HCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
Applications
Biological Information
Purity≥75% by SDS-PAGE
Specific Activity≥270 nmol/min/mg protein
Concentration Label Please refer to vial label for lot-specific concentration
Physicochemical Information
Dimensions
Materials Information
Toxicological Information
Safety Information according to GHS
Safety Information
Product Usage Statements
Storage and Shipping Information
Ship Code Dry Ice Only
Toxicity Standard Handling
Storage ≤ -70°C
Avoid freeze/thaw Avoid freeze/thaw
Do not freeze Ok to freeze
Special InstructionsFollowing reconstitution, aliquot and freeze (-70°C).
Packaging Information
Transport Information
Supplemental Information
Specifications

Documentation

AMPK (α2/β1/γ1), His•Tag®, Human, Recombinant,S. frugiperda Ficha datos de seguridad (MSDS)

Título

Ficha técnica de seguridad del material (MSDS) 

AMPK (α2/β1/γ1), His•Tag®, Human, Recombinant,S. frugiperda Certificados de análisis

CargoNúmero de lote
171539

Referencias bibliográficas

Visión general referencias
Foretz, M., et al. 2005. Diabetes 54, 1331.
Viollet, B., et al. 2003. Biochem. Soc. Trans. 31, 216.
Ficha técnica

Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

Revision15-September-2008 JSW
SynonymsAMP Kinase (α2/β1/γ1)
Application Data
The activity of AMPK (α2/β1/γ1) was measured as outlined in the Recommended Reaction Conditions.
DescriptionFull length, recombinant, human AMPK consisting of subunits α2/β1/γ1, each fused at the C-terminus to a His•Tag® sequence and co-expressed in S. frugiperda insect cells using a baculovirus expression system. AMPK is involved in sensing energy levels in the cell that result from changes in AMP levels. AMPK is an upstream regulator of mTOR activation. The α-subunit contains a kinase domain, the β-subunit contains a glycogen binding domain, and the γ-subunit contains the regulatory AMP binding domain. The active complex is useful in the study of kinase activity regulation and inhibitor screening.
FormLiquid
FormulationIn 150 mM NaCl, 50 mM Tris-HCl, 250 µM DTT, 100 µM EGTA, 100 µM EDTA, 100 µM PMSF, 25% glycerol, pH 7.5.
Concentration Label Please refer to vial label for lot-specific concentration
Recommended reaction conditions
Kinase assay conditions: Dilute purified AMPK in kinase dilution buffer (5 mM MOPS, pH 7.2, 5 mM MgCl2, 2.5 mM β-glycerophosphate, 1 mM EGTA, 400 µM EDTA, 50 ng/µl BSA, 5% glycerol; add DTT to 25 µM just prior to use) to the desired concentration. Mix 10 µl diluted enzyme, 5 µl SAMStide substrate (HMRSAMSGLHLVKRR) (1 mg/ml stock), and 5 µl 500 µM AMP and incubate for 5 min at 30°C. Add 5 µl ATP mixture [(150 µl of 10 mM ATP, 100 µl of 1 mCi/100µl [32P]ATP, and 5.75 ml kinase assay buffer (25 mM MOPS, pH 7.2, 25 mM MgCL2, 12.5 mM β-glycerophosphate, 5 mM EGTA, 2 mM EDTA with 250 µM DTT just prior to use)] and incubate for 15 min at 30°C. Terminate the reaction by spotting 20 µl of the reaction mixture onto a phosphocellulose P81 paper, dry, and wash several times with 1% phosphoric acid prior to counting in the presence of scintillation fluid.
Purity≥75% by SDS-PAGE
Specific activity≥270 nmol/min/mg protein
Unit definitionKinase activity was measured as the molar amount of phosphate incorporated into SAMS peptide substrate (HMRSAMSGLHLVKRR) at 30°C using a final concentration of 50 µM ATP.
Storage ≤ -70°C
Avoid freeze/thaw
Do Not Freeze Ok to freeze
Special InstructionsFollowing reconstitution, aliquot and freeze (-70°C).
Toxicity Standard Handling
ReferencesForetz, M., et al. 2005. Diabetes 54, 1331.
Viollet, B., et al. 2003. Biochem. Soc. Trans. 31, 216.