Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA) (AA 562-724) antibody

Details for Product No. ABIN967707
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Synonyms CLOVE, CWS5, MCAP, MCM, MCMTC, PI3K, p110-alpha, 6330412C24Rik, caPI3K, p110, p110alpha, C-P3K, PI3-kinase, PI3K-alpha, pi3-k, pi3k
AA 562-724
(13), (12), (2), (2), (2), (2), (1), (1), (1), (1)
(79), (42), (37), (14), (12), (2), (1)
(66), (8), (7)
Clonality (Clone)
Monoclonal ()
(2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2)
Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), BioImaging (BI)
(43), (36), (28), (20), (15), (6), (6), (6), (4), (1), (1), (1)
Pubmed 5 references available
Catalog no. ABIN967707
Quantity 50 µg
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Immunogen Human PI3-Kinase alpha subunit
Clone 4
Isotype IgG2a, kappa
Cross-Reactivity Dog (Canine), Rat (Rattus), Mouse (Murine), Chicken
Characteristics 1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to us for technical protocols.
3. This antibody has been developed and certified for the bioimaging application. However, a routine bioimaging test is not performed on every lot. Researchers are encouraged to titrate the reagent for optimal performance.
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. Triton is a trademark of the Dow Chemical Company.
Purification Purified from tissue culture supernatant or ascites by affinity chromatography.
Purity Purified
Alternative Name PI3-Kinase
Background PI3-kinase phosphorylates the D-3 position of the inositol ring of phosphatidylinositol (PtdIns), PtdIns(4)P and PtdIns(4,5)P2 to produce the respective PI3-phosphorylated derivatives. PI3-kinase exists as a heterodimer of 85 kDa (p85) and 110 kDa (p110) subunits. The p85 subunit contains two SH2 domains and an SH3 domain. It associates with and serves as a substrate for activated growth factor receptor tyrosine kinases. p85 may serve as regulator of the catalytic subunit, p110, by acting as the link between PI3-kinase and the ligand-activated receptor. Two distinct forms of the p85 subunit have been described: 1) p85alpha, which binds tightly to the catalytic subunit, and 2) p85ß, a protein whose function is presently unknown. Both isoforms bind to activated receptors and serve as tyrosine kinase substrates.
Molecular Weight 85 kDa
Application Notes Bioimaging
1. Seed the cells in appropriate culture medium at ~10,000 cells per well in an 96-well Imaging Plate and culture overnight.
2. Remove the culture medium from the wells, and fix the cells by adding 100 myl of Fixation Buffer to each well. Incubate for 10 minutes at room temperature (RT).
3. Remove the fixative from the wells, and permeabilize the cells using either 90% methanol, or Triton™ X-100: a. Add 100 myl of -20°C 90% methanol to each well and incubate for 5 minutes at RT. OR b. Add 100 myl of 0.1% Triton™ X-100 to each well and incubate for 5 minutes at RT.
4. Remove the permeabilization buffer, and wash the wells twice with 100 myl of 1× PBS.
5. Remove the PBS, and block the cells by adding 100 myl of to each well. Incubate for 30 minutes at RT.
6. Remove the blocking buffer and add 50 myl of the optimally titrated primary antibody (diluted in Stain Buffer) to each well, and incubate for 1 hour at RT.
7. Remove the primary antibody, and wash the wells three times with 100 myl of 1× PBS.
8. Remove the PBS, and add the second step reagent at its optimally titrated concentration in 50 myl to each well, and incubate in the dark for 1 hour at RT.
9. Remove the second step reagent, and wash the wells three times with 100 myl of 1× PBS.
10. Remove the PBS, and counter-stain the nuclei by adding 200 myl per well of 2 myg/ml Hoechst 33342 in 1× PBS to each well at least 15 minutes before imaging.
11. View and analyze the cells on an appropriate imaging instrument.

Related Products: ABIN967389, ABIN968533

Restrictions For Research Use only
Format Liquid
Concentration 250 µg/ml
Buffer Aqueous buffered solution containing BSA, glycerol.
Preservative Sodium azide
Storage -20 °C
Supplier Images
anti-Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA) (AA 562-724) antibody Western blot analysis of PI3-Kinase on a A431 lysate. Lane 1: 1:2500, lane 2: 1:5000, lane 3: 1:10000 dilution of the PI3-Kinase antibody.
anti-Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA) (AA 562-724) antibody (2) Immunofluorescent staining of A549 (ATCC CCL-185) cells. Cells were seeded in a 96 well imaging plate at ~ 10 000 cells per well. After overnight incubation, cells were stained using the alcohol perm protocol and the anti-PI3-Kinase antibody. The second step reagent was FITC goat anti mouse Ig. The image was taken on a BD Pathway™ 855 Bioimager using a 20x objective. This antibody also stained U-2 OS (ATCC HTB-96) and HeLa (ATCC CCL-2) cells using both the Triton™ X-100 and alcohol perm protocols.
Product cited in: Cantley, Auger, Carpenter et al.: "Oncogenes and signal transduction." in: Cell, Vol. 64, Issue 2, pp. 281-302, 1991 (PubMed).

Kihara, Shimohama, Sawada et al.: "alpha 7 nicotinic receptor transduces signals to phosphatidylinositol 3-kinase to block A beta-amyloid-induced neurotoxicity." in: The Journal of biological chemistry, Vol. 276, Issue 17, pp. 13541-6, 2001 (PubMed).

Zhang, Bontrager, Hemler: "Transmembrane-4 superfamily proteins associate with activated protein kinase C (PKC) and link PKC to specific beta(1) integrins." in: The Journal of biological chemistry, Vol. 276, Issue 27, pp. 25005-13, 2001 (PubMed).

Nguyen, Ho, Beattie et al.: "TEL-JAK2 mediates constitutive activation of the phosphatidylinositol 3'-kinase/protein kinase B signaling pathway." in: The Journal of biological chemistry, Vol. 276, Issue 35, pp. 32704-13, 2001 (PubMed).

Efendiev, Yudowski, Zwiller et al.: "Relevance of dopamine signals anchoring dynamin-2 to the plasma membrane during Na+,K+-ATPase endocytosis." in: The Journal of biological chemistry, Vol. 277, Issue 46, pp. 44108-14, 2002 (PubMed).

Hosts (66), (8), (7)
Reactivities (79), (42), (37), (14), (12), (2), (1)
Applications (43), (36), (28), (20), (15), (6), (6), (6), (4), (1), (1), (1)
Conjugates (2), (2), (2), (2), (2), (2), (2), (2), (2), (2), (2)
Epitopes (13), (12), (2), (2), (2), (2), (1), (1), (1), (1)
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