Analysis Note

ControlNIH-3T3 cell lysate, A431, MCF7 or Raji cells lysate, HeLa cells, breast carcinoma.

Included Positive Antigen Control: Catalog # 12-301, non-stimulated A431 cell lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research CategorySignaling

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).

This Anti-PI3 Kinase Antibody, p85, N-SH3, clone AB6 is validated for use in IC, IP, WB for the detection of PI3 Kinase.

Research Sub CategoryPI3K, Akt, & mTOR SignalingKinases & Phosphatases

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.

General description

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.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-403

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Purified mouse monoclonal IgG1κ in buffer containing PBS, pH 7.2 with 50% glycerol.

Protein A chromatography

Format: Purified

Quality

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.

Specificity

This antibody recognizes p85 alpha subunit of PI3 Kinase; does not recognize p85 beta subunit.

Storage and Stability

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 -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Target description

85 kDa