Recombinant AKT3 protein

Catalog No: 81147 Format: 20 µg $420 Buy
Catalog No: 81847 Format: 1 mg $3,550 Buy

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Expressed In: Baculovirus Protein Species: Human

Contents

A representative Technical Data Sheet (TDS) is provided here. Please refer to the lot-specific TDS you will receive with your order for the lot-specific buffer contents and protein concentration.

Background

AKT3 (AKT Serine/Threonine Kinase 3), also known as Protein Kinase B Gamma or PRKBG, is one of 3 closely related serine/threonine-protein kinases (AKT3, AKT3 and AKT3) called the AKT kinase, and which regulate many processes including metabolism, proliferation, cell survival, growth and angiogenesis. This is mediated through serine and/or threonine phosphorylation of a range of downstream substrates. Over 100 substrate candidates have been reported so far, but for most of them, no isoform specificity has been reported. AKT3 is the least studied AKT isoform. It plays an important role in brain development and is crucial for the viability of malignant glioma cells. AKT3 isoform may also be the key molecule in up-regulation and down-regulation of MMP13 via IL13. It is required for the coordination of mitochondrial biogenesis with growth factor-induced increases in cellular energy demands. Down-regulation by RNA interference reduces the expression of the phosphorylated form of BAD, resulting in the induction of caspase-dependent apoptosis.

Application Notes

Recombinant AKT3 protein is suitable for use in the study of enzyme kinetics, inhibitor screening, and selectivity profiling.

Kinase Activity Assay Conditions: 1 µM STK S3 substrate was incubated with different concentrations AKT3 protein in a 10 µl reaction system containing 1×Enzymatic Buffer, 5 mM MgCl2, 1 mM DTT, 5 nM SEB and 100 µM ATP for 1 hour. The 10 µl detection reagents containing STK antibody and SA-XL665, each of which was 1:100 diluted with 1× Detection Buffer were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature, and HTRF KinEASE STK assay was used to detect the enzymatic activity.

Protein Details

Recombinant AKT3 protein was expressed in a baculovirus expression system as the full length protein (accession number NP_005456.1) with an N-terminal 6XHis and FLAG-Tag. The molecular weight of the protein is 60.8 kDa.

 

Recombinant AKT3 protein gel
10% SDS-PAGE Coomassie staining
MW: 60.8 kDa
Purity: ≥90%

HTRF assay for recombinant AKT3 protein activity
1 µM STK S3 substrate was incubated with different concentrations AKT3 protein in a 10 µl reaction system for 1 hour. The 10 µl detection reagents were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature, and HTRF KinEASE STK assay was used to detect the enzymatic activity.

HTRF assay for recombinant AKT3 protein activity
1 µM STK S3 substrate was incubated with different concentrations AKT3 protein in a 10 µl reaction system for 1 hour. The 10 µl detection reagents were added and incubated with the reactions for 30 min. All the operations and reactions were performed at room temperature, and HTRF KinEASE STK assay was used to detect the enzymatic activity.

Storage

Recombinant proteins in solution are temperature sensitive and must be stored at -80°C to prevent degradation. Avoid repeated freeze/thaw cycles and keep on ice when not in storage.

Guarantee

This product is for research use only and is not for use in diagnostic procedures. This product is guaranteed for 6 months from date of arrival.