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Encompassing Amino Acids: full length
Applications: Useful for the study of enzyme kinetics, screening inhibitors, and selectivity profiling.
Assay Conditions: Assay was done in Kinase buffer containing 1 mM DTT using AMARApeptide as a substrate (0.1 mg/ml), 20 mM ATP and 100 µM AMP . Reaction was done at 30°C for 45 min. Amount of ATP transferred was calculated using Kinase-Glo reagent (Promega)
Background: AMPK is a heterotrimer protein kinase consisting of an α catalytic subunit, and non-catalytic β and γ subunits. AMPK is an important energy-sensing enzyme that monitors cellular energy status (1). In response to cellular metabolic stresses, AMPK is activated and phosphorylates and inactivates acetyl-CoA carboxylase (ACC) and beta-hydroxy beta-methylglutaryl-CoA reductase (HMGCR), key enzymes involved in regulating biosynthesis of fatty acid and cholesterol (2).
Description: Recombinant full-lengthhuman AMPK (combination of A1/B1/G1subunits) was expressed by baculovirus inSf9 insect cells using a C-terminal His tag.The gene accession number for the threesubunits (A1/B1/G1) is NM_006251,NM_006253, and NM_002733,respectively. This protein has beenactivated with CAMKK.
Format: Aqueous buffer solution
Formulation: 50 mM sodium phosphate, pH 7.0, 300 mM NaCl, 150 mM imidazole, 0.1 mM PMSF, 0.25 mM DTT, 25% glycerol.
Genbank: NM_006251(A1), NM_006253(B1), NM_002733 (G1)
Host Cell Line: Sf9 cells
Purity: ≥95%
Storage Stability: At least 6 months at -80°C.
Tags: C-terminal His-tags
Uniprot: A1: Q13131; B1: Q9Y478; G1: P54619
Warnings: Avoid freeze/thaw cycles. Storing diluted enzyme is not recommended. If necessary, use carrier protein (BSA 0.1 - 0.5%).
Biosafety Level: Not applicable (BSL-1)
References: 1. Minokoshi, Y. et al. Nature 428: 569-574, 2004.
2. Hardie, D G. et al. Eur J Biochem. 1997 Jun 1,246(2):259-73.
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