PIK3CA antibody (AA 562-724)

Catalog No. ABIN967707

Product Details anti-PIK3CA Antibody

Target: PIK3CA (Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA))

Binding Specificity: AA 562-724

Reactivity: Human, Mouse, Rat, Dog, Chicken

Host: Mouse

Clonality: Monoclonal

Conjugate: This PIK3CA antibody is un-conjugated

Application: Western Blotting (WB), Immunohistochemistry (IHC), Immunoprecipitation (IP), BioImaging (BI)

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: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Human PI3-Kinase alpha subunit aa. 562-724

Clone: 4-PI3

Isotype: IgG2a

Application Details

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.

Comment:

Related Products: ABIN967389, ABIN968533

Restrictions: For Research Use only

Handling

Format: Liquid

Concentration: 250 μg/mL

Buffer: Aqueous buffered solution containing BSA, glycerol, and ≤0.09 % sodium azide.

Preservative: Sodium azide

Precaution of Use: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Storage: -20 °C

Storage Comment: Store undiluted at -20°C.

References for anti-PIK3CA antibody (ABIN967707)

Efendiev, Yudowski, Zwiller, Leibiger, Katz, Berggren, Pedemonte, Leibiger, Bertorello: "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).

Kihara, Shimohama, Sawada, Honda, Nakamizo, Shibasaki, Kume, Akaike: "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).

Nguyen, Ho, Beattie, Barber: "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).

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).

Cantley, Auger, Carpenter, Duckworth, Graziani, Kapeller, Soltoff: "Oncogenes and signal transduction." in: Cell, Vol. 64, Issue 2, pp. 281-302, (1991) (PubMed).

Target Details for PIK3CA

Target: PIK3CA (Phosphoinositide-3-Kinase, Catalytic, alpha Polypeptide (PIK3CA))

Alternative Name: PI3-Kinase (PIK3CA Products)

Synonyms: CLOVE antibody, CWS5 antibody, MCAP antibody, MCM antibody, MCMTC antibody, PI3K antibody, p110-alpha antibody, C-P3K antibody, PI3-kinase antibody, PI3K-alpha antibody, pi3-k antibody, pi3k antibody, AA414921 antibody, C530050K14 antibody, p50alpha antibody, p55alpha antibody, p85alpha antibody, 6330412C24Rik antibody, caPI3K antibody, p110 antibody, p110alpha antibody, phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha antibody, Phosphoinositide-3-kinase, catalytic, alpha polypeptide antibody, phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha antibody, phosphoinositide-3-kinase regulatory subunit 1 antibody, PIK3CA antibody, GL50803_14855 antibody, pik3ca antibody, Pik3ca antibody, Pik3r1 antibody

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

Pathways: PI3K-Akt Signaling, RTK Signaling, TCR Signaling, AMPK Signaling, Interferon-gamma Pathway, TLR Signaling, Fc-epsilon Receptor Signaling Pathway, EGFR Signaling Pathway, Neurotrophin Signaling Pathway, Inositol Metabolic Process, Hepatitis C, CXCR4-mediated Signaling Events, Signaling Events mediated by VEGFR1 and VEGFR2, Signaling of Hepatocyte Growth Factor Receptor, VEGFR1 Specific Signals, VEGF Signaling