Product Pathways - PI3K / Akt Signaling
PhosphoPlus® Akt Activation Kit #9280
|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|Phospho-Akt (Thr308) (C31E5E) Rabbit mAb #2965||100 µl||W IF-IC F||H M R Hm Mk||60||Rabbit IgG|
|Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060||100 µl||W IP IHC-P IHC-F IF-IC F||H M R Hm Mk Dm Z B (C) (X) (Dg) (Pg)||60||Rabbit IgG|
|Akt (pan) (C67E7) Rabbit mAb #4691||100 µl||W IP IHC-P IF-IC F||H M R Mk Dm (Pg)||60||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
|Anti-biotin, HRP-linked Antibody #7075||100 µl||Goat|
|Biotinylated Protein Ladder Detection Pack #7727||100 µl|
|20X LumiGLO® Reagent and 20X Peroxide #7003||5 ml each|
Reactivity Key: H=Human M=Mouse R=Rat Hm=Hamster Mk=Monkey C=Chicken Dm=D. melanogaster X=Xenopus Z=Zebrafish B=Bovine Dg=Dog Pg=Pig
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Specificity / Sensitivity
The phospho-Akt antibodies detect endogenous levels of Akt only when phosphorylated at either Ser473 or Thr308. The Akt antibody detects endogenous levels of total Akt1, Akt2 and Akt3 proteins independent of their phosphorylation state. It does not cross-react with related kinases.
Western blot analysis of extracts from NIH/3T3 and Jurkat cells, untreated, PDGF-treated or LY294002-treated as indicated, using Phospho-Akt (Thr308) (C31E5) Rabbit mAb #2965 (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Western blot analysis of extracts from PC3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum- starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060.
Flow cytometric analysis of Jurkat cells, untreated (green) or LY294002, Wortmannin and U0126-treated (blue) using Phospho-Akt (Thr308) (C31E5) Rabbit mAb #2965.
Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002, wortmannin and U0126 (blue), using Phospho-Akt (Ser473) (D9E) Rabbit mAb #4060 compared to a nonspecific negative control antibody (red).
Kit includes:#2965 (100uL)#4060 (100uL)#4961 (100uL)#7074 (100uL)#7075 100ul7003 5 ml7727 100ul
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding either Ser473, Thr308 or from the carboxy-terminal sequence of mouse Akt.
Akt, also referred to as PKB or Rac, plays a critical role in controlling survival and apoptosis (1-3). This protein kinase is activated by insulin and various growth and survival factors to function in a wortmannin-sensitive pathway involving PI3 kinase (2,3). Akt is activated by phospholipid binding and activation loop phosphorylation at Thr308 by PDK1 (4) and by phosphorylation within the carboxy terminus at Ser473. The previously elusive PDK2 responsible for phosphorylation of Akt at Ser473 has been identified as mammalian target of rapamycin (mTOR) in a rapamycin-insensitive complex with rictor and Sin1 (5,6). Akt promotes cell survival by inhibiting apoptosis through phosphorylation and inactivation of several targets, including Bad (7), forkhead transcription factors (8), c-Raf (9), and caspase-9. PTEN phosphatase is a major negative regulator of the PI3 kinase/Akt signaling pathway (10). LY294002 is a specific PI3 kinase inhibitor (11). Another essential Akt function is the regulation of glycogen synthesis through phosphorylation and inactivation of GSK-3α and β (12,13). Akt may also play a role in insulin stimulation of glucose transport (12). In addition to its role in survival and glycogen synthesis, Akt is involved in cell cycle regulation by preventing GSK-3β-mediated phosphorylation and degradation of cyclin D1 (14) and by negatively regulating the cyclin dependent kinase inhibitors p27 Kip1 (15) and p21 Waf1/Cip1 (16). Akt also plays a critical role in cell growth by directly phosphorylating mTOR in a rapamycin-sensitive complex containing raptor (17). More importantly, Akt phosphorylates and inactivates tuberin (TSC2), an inhibitor of mTOR within the mTOR-raptor complex (18,19).
The Akt activation kit provides an economical means to evaluate the activation status of Akt at both the Ser473 and Thr308 positions as well as the total Akt protein level. The kit includes enough primary and secondary antibodies to perform ten Western mini-blot experiments.
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* Product-specific protocol.
- 4058 Phospho-Akt (Ser473) (193H12) Rabbit mAb
- 4056 Phospho-Akt (Thr308) (244F9) Rabbit mAb
- 2938 Akt1 (C73H10) Rabbit mAb
- 9916 Phospho-Akt Pathway Antibody Sampler Kit
- 9940 Akt Isoform Antibody Sampler Kit
- 7160 PathScan® Phospho-Akt1 (Ser473) Sandwich ELISA Kit
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 3063 Akt2 (D6G4) Rabbit mAb
- 8018 Akt3 (L47B1) Mouse mAb
For Research Use Only. Not For Use In Diagnostic Procedures.