Description
General description
85 kDa
The enzyme phosphatidylinositol 3 kinase (PI3 kinase) is a lipid kinase that is thought to phosphorylate second messengers in growth signaling pathways. PI3K exists as a heterodimer of catalytic 110 kDa (p110) and regulatory 85 kDa (p85, p50 or p55) subunits. The p85 unit consists of two closely related proteins: p85 alpha, which binds tightly to the catalytic subunit and p85 beta, a protein of unknown function. They share 62% amino acid sequence identity. p85 alpha lacks PI3 kinase activity, but may serve as regulator of the catalytic subunit, p110, by acting as the link between PI3K and the ligand activated receptor. Both isoforms bind to activated receptors and serve as tyrosine kinase substrates.
Immunogen
Recombinant human p85 alpha PI3 Kinase expressed in E. Coli. Clone AB6.
Application
Immunocytochemistry:
10 µg/mL of a previous lot of this antibody was used in immunocytochemistry.
Immunoprecipitation:
4 µg of a previous lot immunoprecipitated PI3 Kinase from 500 µg of an A431 RIPA lysate, as confirmed by immunoblot using a polyclonal PI3 Kinase antibody (Catalog # 06-195).
Research Category
Signaling
Research Sub Category
PI3K, Akt, & mTOR Signaling
Kinases & Phosphatases
This Anti-PI3 Kinase Antibody, p85, N-SH3, clone AB6 is validated for use in IC, IP, WB for the detection of PI3 Kinase.
Biochem/physiol Actions
This antibody recognizes p85 alpha subunit of PI3 Kinase; does not recognize p85 beta subunit.
Physical form
Format: Purified
Protein A chromatography
Purified mouse monoclonal IgG1κ in buffer containing PBS, pH 7.2 with 50% glycerol.
Preparation Note
Stable for 1 year at -20°C from date of receipt.
Handling Recommendations:
Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20ºC. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20ºC may cause glycerol-containing solutions to become frozen during storage.
Analysis Note
Control
Positive Antigen Control: Catalog #12-301, non-stimulated A431 cell lysate. Add 2.5µL of 2-mercaptoethanol/100µL of lysate and boil for 5 minutes to reduce the preparation. Load 20µg of reduced lysate per lane for minigels.
Routinely evaluated by Western Blot on RIPA lysates from human A431 cells, or RIPA lysates from rat L6 myoblasts.
Western Blot Analysis:
0.5-2 µg/mL of this lot detected PI3 Kinase in RIPA lysates from human A431 cells; previous lots detected PI3 Kinase in RIPA lysates from rat L6 myoblasts.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Replaces: 04-403
Legal Information
UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
