Recombinant Human AMPK alpha 1 Protein

Beta LifeScience SKU/CAT #: BLPSN-0183

Recombinant Human AMPK alpha 1 Protein

Beta LifeScience SKU/CAT #: BLPSN-0183
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Product Overview

Tag N/A
Host Species Human
Accession P54619
Synonym AMPK, AMPKa1
Background The PRKAA1 gene encodes the catalytic alpha-subunit of 5' AMP-activated protein kinase (AMPK). AMPK is a cellular energy sensor that maintains energy homeostasis within the cell and is activated when the AMP/ATP ratio increases. Moreover, AMPK has been shown to be involved in inhibiting tumour growth and metastasis, and has also been implicated in the pathology of neurodegenerative and cardiac disorders. All 5 tested tag SNPs of the PRKAA1 gene (rs1336177, rs154268, rs385486, rs688293, and rs174991) were significantly associated with gastric cancer.An inhibitor of PRKAA1 promoted the secretion of IL6, whereas an agonist of PRKAA1 suppressed the production of IL6.
Description A DNA sequence encoding the human PRKAG1 (P54619) (Met 1-Pro 331), constructed the plasmid 1; A DNA sequence encoding the human PRKAB1 (Q9Y478) (Met 1-Ile 270), constructed the plasmid 2; A DNA sequence encoding the human PRKAA2 (P54646) (Met 1-Arg 552) was fused with the N-terminal His-tagged GST tag at the N-terminus, constructed the plasmid 3. The three plasmids were co-expressed and the heterotrimer was purified.
Source Baculovirus-Insect Cells
Predicted N Terminal Met & Met & His
AA Sequence Met 1-Pro 331
Molecular Weight The recombinant heterotrimer of human AMPK (PRKAG1 / PRKAB1 / PRKAA2) has a calculated molecular mass of 158 (38+30+90) KDa. The apparent molecular mass is approximately 40, 42 & 95 KDa respectively in SDS-PAGE under reducing conditions.
Purity >94% as determined by SDS-PAGE
Endotoxin < 1.0 EU per μg of the protein as determined by the LAL method
Bioactivity The specific activity was determined to be 4 nmol/min/mg using synthetic SAMS peptide (HMRSAMSGLHLVKRR) as substrate.
Formulation Supplied as sterile 50mM Tris, 200mM NaCl, 1mM EDTA, 1mM DTT, 0.5mM PMSF, 10% gly, 1mM GSH, pH 7.4.
Stability The recombinant proteins are stable for up to 1 year from date of receipt at -70°C.
Usage For Research Use Only
Storage Store the protein under sterile conditions at -20°C to -80°C. It is recommended that the protein be aliquoted for optimal storage. Avoid repeated freeze-thaw cycles.

FAQs

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Proteins are sensitive to heat, and freeze-drying can preserve the activity of the majority of proteins. It improves protein stability, extends storage time, and reduces shipping costs. However, freeze-drying can also lead to the loss of the active portion of the protein and cause aggregation and denaturation issues. Nonetheless, these adverse effects can be minimized by incorporating protective agents such as stabilizers, additives, and excipients, and by carefully controlling various lyophilization conditions.

Commonly used protectant include saccharides, polyols, polymers, surfactants, some proteins and amino acids etc. We usually add 8% (mass ratio by volume) of trehalose and mannitol as lyoprotectant. Trehalose can significantly prevent the alter of the protein secondary structure, the extension and aggregation of proteins during freeze-drying process; mannitol is also a universal applied protectant and fillers, which can reduce the aggregation of certain proteins after lyophilization.

Our protein products do not contain carrier protein or other additives (such as bovine serum albumin (BSA), human serum albumin (HSA) and sucrose, etc., and when lyophilized with the solution with the lowest salt content, they often cannot form A white grid structure, but a small amount of protein is deposited in the tube during the freeze-drying process, forming a thin or invisible transparent protein layer.

Reminder: Before opening the tube cap, we recommend that you quickly centrifuge for 20-30 seconds in a small centrifuge, so that the protein attached to the tube cap or the tube wall can be aggregated at the bottom of the tube. Our quality control procedures ensure that each tube contains the correct amount of protein, and although sometimes you can't see the protein powder, the amount of protein in the tube is still very precise.

To learn more about how to properly dissolve the lyophilized recombinant protein, please visit Lyophilization FAQs.

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