Recombinant PI3Kα (G106V) / p85 Complex

Catalog No: 82179 Format: 20 µg $480 Add to Cart
Catalog No: 82779 Format: 1 mg $4,150 Add to Cart

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Applications

Application Notes

This product was manufactured as described in Protein Details. Where possible, Active Motif has developed functional or activity assays for recombinant proteins. Additional characterization such as enzyme kinetic activity assays, inhibitor screening or other biological activity assays may not have been performed for every product.  All available data for this product is shown. 

References

Human

 
Recombinant PI3Kα protein gel

Recombinant PI3Kα (G106V / p85) protein gel
10% SDS-PAGE gel
MW: PI3Kα (G106V): 125.6 kDa
p85: 83.6 kDa
Purity: >90%

ADP-Glo Assay for PI3Kα activity.

ADP-Glo assay for PI3Kα (G106V) / p85 Complex activity
0.125 µg/µl PI-3 Kinase Lipid Substrate was incubated with different concentrations of PI3Kα (G106V) / p85 Complex protein in a 10 µl reaction system containing 40 mM Tris-HCl pH 7.4, 20 mM MgCl2, 50 µM DTT, 0.1 mg/ml BSA and 100 µM ATP for 1 hour, then add 10 µl ADP-Glo Reagent at RT for 60min.Then add 20 µl Kinase Detection Reagent incubates at RT for 60min. All the operations and reactions were performed at RT. Finally, Luminescence measurement is collected by BMG

ADP-Glo Assay for PI3Kα activity.

ADP-Glo assay for PI3Kα (G106V) / p85 Complex activity
0.125 µg/µl PI-3 Kinase Lipid Substrate was incubated with different concentrations of PI3Kα (G106V) / p85 Complex protein in a 10 µl reaction system containing 40 mM Tris-HCl pH 7.4, 20 mM MgCl2, 50 µM DTT, 0.1 mg/ml BSA and 100 µM ATP for 1 hour, then add 10 µl ADP-Glo Reagent at RT for 60min.Then add 20 µl Kinase Detection Reagent incubates at RT for 60min. All the operations and reactions were performed at RT. Finally, Luminescence measurement is collected by BMG

Background

Phosphoinositide 3-kinase (PI3K) family of lipid kinases are divided into three major classes based on primary sequence, substrate preference, and regulation. While class IA PI3Ks are heterodimeric enzymes composed of a catalytic subunit (p110a, p110b, or p110d) complexed with one of five regulatory subunits (p85a, p55a, p50a, p85b, or p55g), the class IB enzyme is a dimer made of p110g catalytic subunit and p101or p84 regulatory subunit. The class I catalytic subunit polypeptides p110a, p110b, p110d, and p110g are encoded by PIK3CA, PIK3CB, PIK3CD, and PIK3CG, respectively. The regulatory subunits are encoded by five genes: PIK3R1 codes p85a, p55a, and p50a; PIK3R2 codes p85b; PIK3R3 codes p55g; PIK3R5 codes p101; and PIK3R6 codes p84. PI3K is a lipid and protein kinase that can be activated by receptor tyrosine kinases, cytokine receptors, chemokine G protein–coupled receptors, or oncogenes, including RAS. PI3K converts phosphatidylinositol 4,5-bisphosphate (PIP2) to phosphatidylinositol-3,4,5-triphosphate (PIP3), which leads to activation of pyruvate dehydrogenase kinase 1 (PDK1) and the serine/threonine kinase AKT (PKB) (7). AKT has many downstream effectors, which regulate the cell cycle, apoptosis, protein translation, autophagy, and metabolism. Most cytokines and RTK ligands signal through class IA PI3K.

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.

Application Key

  • ChIP = Chromatin Immunoprecipitation;
  • ChIP = ChIP Sequencing;
  • CUT&RUN = Cleavage Under Targets and Release Using Nuclease;
  • CUT&Tag = Cleavage Under Targets and Tagmentation;
  • DB = Dot Blot;
  • ELISA = Enzyme-linked Immunosorbent Assay;
  • EMSA = Electrophoretic Mobility Shift Assay
  • FC = Flow Cytometry;
  • ICC = Immunocytochemistry;
  • IF = Immunofluorescence;
  • IHC = Immunohistochemistry;
  • IP = Immunoprecipitation;
  • MeDIP = Methyl-DNA Immunoprecipitation;
  • NEU = Neutralizing;
  • TIP-ChIP = Tagmented, Indexed, and Pooled Chromatin Immunoprecipitation;
  • WB = Western Blotting