Western blot analysis of extracts from various cell types using Akt1 (C73H10) Rabbit mAb.
Western blot analysis of extracts from various cell lines using Akt2 (D6G4) Rabbit mAb (upper) and Akt1 (C73H10) Rabbit mAb #2938 (lower).
Western blot analysis of extracts from various cell lines using Akt3 (L47B1) Mouse mAb (upper) and Akt (pan) (40D4) Mouse mAb #2920 (lower).
Flow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
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.
Western blot analysis of extracts from Akt1 (lane 1), Akt2 (lane 2) and Akt3 (lane 3) knock-out mouse embryonic fibroblasts (MEFs) and matched wild-type MEFs (lane 4) using Akt1 (C73H10) Rabbit mAb (upper) and Akt2 Antibody #2962 (lower).
Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Akt (pan) (C67E7) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).
Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins using Akt1 (C73H10) Rabbit mAb (upper) and Akt (pan) (11E7) Rabbit mAb #4685 (lower).
Immunohistochemical analysis of paraffin-embedded human melanoma using Akt (pan) (C67E7) Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Akt (pan) (C67E7) Rabbit mAb in the presence of control peptide (left) or Akt (pan) Blocking Peptide #1085 (right).
Immunohistochemical analysis using Akt (pan) (C67E7) Rabbit mAb on SignalSlide (TM) Phospho-Akt (Ser473) IHC Controls #8101 (paraffin-embedded LNCaP cells, untreated (left) or LY294002-treated (right)).
Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins, and extracts from various cell lines, using Akt (pan) (C67E7) Rabbit mAb.
|Akt1 (C73H10) Rabbit mAb 2938||20 µl||
||H M R Mk||60||Rabbit IgG|
|Akt2 (D6G4) Rabbit mAb 3063||20 µl||
||H M R Mk||60||Rabbit IgG|
|Akt3 (L47B1) Mouse mAb 8018||20 µl||
||H M R Hm||60||Mouse IgG1|
|Akt (pan) (C67E7) Rabbit mAb 4691||20 µl||
||H M R Mk Dm||60||Rabbit IgG|
|Anti-rabbit IgG, HRP-linked Antibody 7074||100 µl||
Akt Isoform Specific Antibody Sampler Kit provides an economical means of distinguishing between the three Akt isoforms, Akt1, Akt2 and Akt3, as well as a pan-Akt antibody. The kit provides enough primary and secondary antibodies to perform two Western blot experiments.
Each isoform-specific antibody recognizes endogenous levels of the specified Akt isoform independent of its phosphorylation state. Neither antibody cross-reacts with the two other isoforms. Akt Antibody recognizes endogenous levels of all three Akt isoforms.
Each isoform-specific antibody is produced by immunizing mice or rabbits with a synthetic peptide that is specific for the Akt isoform. Akt1 Monoclonal antibody is produced by immunizing animals with a synthetic peptide surrounding Leu110 of human Akt1. Akt2 monoclonal antibody #3063 is produced by immunizing rabbits with a synthetic peptide corresponding to residues of human Akt2. Akt3 monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg116 of human Akt3 protein. Akt(pan) monoclonal antibody is produced by immunizing animals with a synthetic peptide at 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).
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