Western blot analysis of extracts from NIH/3T3-Src cells, untreated or treated with lambda phosphatase and from C2C12 cells, untreated or treated with H2O2, using Phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) Antibody.
Western blot analysis of extracts from Jurkat and NIH/3T3 cells, using PI3 Kinase p85 (19H8) Rabbit mAb.
Western blot analysis of extracts from HeLa cells and neonatal mouse brain using PI3 Kinase p110α (C73F8) Rabbit mAb.
Western blot analysis of extracts from MCF-7 and K-562 cells using PI3 Kinase p110β (C33D4) Rabbit mAb.
Western blot analysis of extracts from 293 and RN33B cells using PI3 Kinase Class III (D4E2) Rabbit mAb.
Western blot analysis of extracts from NIH/3T3, Jurkat and K562 cells using PI3 Kinase p110γ (D55D5) Rabbit mAb.
After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.
|Phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) Antibody 4228||20 µl||
||M||60 and 85||Rabbit|
|PI3 Kinase p85 (19H8) Rabbit mAb 4257||20 µl||
||H M R||85||Rabbit IgG|
|PI3 Kinase p110α (C73F8) Rabbit mAb 4249||20 µl||
||H M R B||110||Rabbit IgG|
|PI3 Kinase p110β (C33D4) Rabbit mAb 3011||20 µl||
|PI3 Kinase Class III (D4E2) Rabbit mAb 3358||20 µl||
||H M R Mk||100||Rabbit IgG|
|PI3 Kinase p110γ (D55D5) Rabbit mAb 5405||20 µl||
||H M||110||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
The PI3 Kinase Sampler Kit provides an economical means of studying PI3 kinase subunits in cells. The kit contains enough primary and secondary antibodies to perform two Western blot experiments per primary antibody.
Each antibody in the PI3 Kinase Antibody Sampler Kit recognizes only its specific target and does not cross-react with other family members. Each total antibody detects endogenous levels of its target protein. Phospho-PI3K p85 (Tyr458)/p55 (Tyr199) Antibody detects endogenous levels of p85/p55 only when phosphorylated at Tyr458/Tyr199.
Phospho-specific polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Tyr458 of mouse p85. Polyclonal antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibodies are produced by immunizing animals with synthetic peptides corresponding to the sequence of human PI3K p85, corresponding to human PI3K p110-gamma, surrounding Asp520 from human PI3K p110α, containing human PI3K p110β, and the amino-terminal sequence of human PI3K class III.
Phosphoinositide 3-kinase (PI3K) catalyzes the production of phosphatidylinositol-3,4,5-triphosphate by phosphorylating phosphatidylinositol (PI), phosphatidylinositol-4-phosphate (PIP), and phosphatidylinositol-4,5-bisphosphate (PIP2). Growth factors and hormones trigger this phosphorylation event, which in turn coordinates cell growth, cell cycle entry, cell migration, and cell survival (1). PTEN reverses this process, and research studies have shown that the PI3K signaling pathway is constitutively activated in human cancers that have loss of function of PTEN (2). PI3Ks are composed of a catalytic subunit (p110) and a regulatory subunit. Various isoforms of the catalytic subunit (p110α, p110β, p110γ, and p110δ) have been isolated, and the regulatory subunits that associate with p110α, p110β, and p110δ are p85α and p85β (3). In contrast, p110γ associates with a p101 regulatory subunit that is unrelated to p85. Furthermore, p110γ is activated by βγ subunits of heterotrimeric G proteins (4).
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