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Flow cytometric analysis of Jurkat cells, untreated (green) or treated with LY294002 #9901, Wortmannin #9951, and U0126 #9903 (50 μM, 1 μM, and 10 μM, 2 hr; blue) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (dashed lines). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.
Flow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Flow cytometric analysis of Akt1 (-/-) mouse embryonic fibroblast (MEF) cells (blue) or Akt2 (-/-) MEF cells (green), PDGF-treated (100 ng/mL, 15 min), using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary Ab.
Western blot analysis of extracts from various cell types using Akt1 (C73H10) Rabbit mAb.
Western blot analysis of extracts from Akt1 (-/-) mouse embryonic fibroblasts (MEF) or Akt2 (-/-) MEF, untreated (-) or treated with Human Platelet-Derived Growth Factor AA (hPDGF-AA) #8913 (100 ng/ml, 15 min; +), using Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle), or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Western blot analysis of extracts from various cell lines using Akt2 (D6G4) Rabbit mAb (upper) and Akt1 (C73H10) Rabbit mAb #2938 (lower).
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.
Confocal immunofluorescent analysis of C2C12 cells, LY294002-treated (left) or insulin-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin #8953 (red). Blue pseudocolor = DRAQ5®#4084 (fluorescent DNA dye).
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).
Confocal immunofluorescent analysis of Akt1 (-/-) mouse embryonic fibroblast (MEF) (left column), or Akt2 (-/-) MEF (right column) cells, stimulated with hPDGF #8913 (100 ng/ml, 15 min) (top row) or inhibited with LY294002 #9901 (bottom row), using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
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).
Western blot analysis of extracts from LNCaP cells, transfected with a construct expressing Akt1 shRNA or Akt2 shRNA, uninduced (-) or doxycycline-induced for the indicated times, using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) #9018 (top), Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) (2nd from top), Akt1 (C73H10) Rabbit mAb #2938 (middle), Akt2 (D6G4) Rabbit mAb #3063 (2nd from bottom), or PI3 Kinase p85 (19H8) Rabbit mAb #4257 (bottom). (Data kindly provided by Drs. Rebecca Chin and Alex Toker, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA).
Immunohistochemical analysis of paraffin-embedded MDA-MB-468 xenograft using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (left) or PTEN (138G6) Rabbit mAb #9559 (right). Note the presence of P-Akt staining in the PTEN deficient MDA-MB-468 cells.
Immunohistochemical analysis of paraffin-embedded human melanoma using Akt (pan) (C67E7) Rabbit mAb.
Western blot analysis of extracts from LNCaP cells, transfected with a construct expressing Akt1 shRNA or Akt2 shRNA, uninduced (-) or doxycycline-induced for the indicated times, using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (top), Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) #8599 (2nd from top), Akt1 (C73H10) Rabbit mAb #2938 (middle), Akt2 (D6G4) Rabbit mAb #3063 (2nd from bottom), or PI3 Kinase p85 (19H8) Rabbit mAb #4257 (bottom). (Data kindly provided by Drs. Rebecca Chin and Alex Toker, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA).
Western blot analysis of recombinant Akt1, Akt2 and Akt3 proteins using Akt1 (C73H10) Rabbit mAb (upper) and Akt (pan) (11E7) Rabbit mAb #4685 (lower).
Western blot analysis of purified recombinant phospho-Akt1, phospho-Akt2 and phospho-Akt3 proteins using Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma comparing SignalStain® Antibody Diluent #8112 (left) to TBST/5% normal goat serum (right) using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060.
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).
Western blot analysis of purified recombinant phospho-Akt1, phospho-Akt2 and phospho-Akt3 proteins using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle) and Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
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 extracts from Akt1 (-/-) mouse embryonic fibroblast (MEF) or Akt2 (-/-) MEF, untreated or stimulated with hPDGF #8913 (100 ng/ml, 15 min), using Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) (upper), Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb #4060 (middle), or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
Immunohistochemical analysis using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb on SignalSlide® 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.
Immunohistochemical analysis of paraffin-embedded human lung carcinoma using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
Immunohistochemical analysis of paraffin-embedded PTEN heterozygous mutant mouse endometrium using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb. (Tissue section courtesy of Dr. Sabina Signoretti, Brigham and Women's Hospital, Harvard Medical School, Boston, MA.)
Immunohistochemical analysis of paraffin-embedded U-87MG xenograft, untreated (left) or lambda phosphatase-treated (right), using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb.
Western blot analysis of extracts from PC-3 cells, untreated or LY294002/wortmannin-treated, and NIH/3T3 cells, serum-starved or PDGF-treated, using Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (upper) or Akt (pan) (C67E7) Rabbit mAb #4691 (lower).
| 製品番号 | サイズ | 価格 | 在庫 |
|---|---|---|---|
| 10396T | 1 Kit (6 x 20 µl) |
| Product Includes | Quantity | Applications | Reactivity | MW(kDa) | Isotype |
|---|---|---|---|---|---|
| Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb 4060 | 20 µl |
|
H M R Hm Mk Dm Z B | 60 | Rabbit IgG |
| Akt (pan) (C67E7) Rabbit mAb 4691 | 20 µl |
|
H M R Mk Dm | 60 | Rabbit IgG |
| Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) 9018 | 20 µl |
|
H M R | 60 | Rabbit IgG |
| Akt1 (C73H10) Rabbit mAb 2938 | 20 µl |
|
H M R Mk | 60 | Rabbit IgG |
| Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) 8599 | 20 µl |
|
H M R | 60 | Rabbit IgG |
| Akt2 (D6G4) Rabbit mAb 3063 | 20 µl |
|
H M R Mk | 60 | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody 7074 | 100 µl |
|
Goat |
Product Information
The Phospho-Akt Isoform Antibody Sampler Kit provides an economical means of detecting the activation of Akt family members using phospho-specific and control antibodies. The kit contains enough primary antibodies to perform at least two western blot experiments per antibody.
Each antibody in this kit recognizes endogenous levels of its specific target protein. Akt (pan) (C67E7) Rabbit mAb does not cross-react with other related proteins. Akt1 (C73H10) Rabbit mAb does not cross-react with Akt2 or Akt3. Akt2 (D6G4) Rabbit mAb does not cross-react with Akt1 or Akt3. Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb detects endogenous levels of Akt only when phosphorylated at Ser473. Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) recognizes endogenous levels of Akt1 protein only when phosphorylated at Ser473. It does not detect Akt2 protein when phosphorylated at Ser474. Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) recognizes endogenous levels of Akt2 protein only when phosphorylated at Ser474. This antibody does not cross-react with Akt1 protein when phosphorylated at Ser473 or with Akt3 protein when phosphorylated at Ser472.
Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues around Ser473 of human Akt. Akt (pan) (C67E7) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues in the carboxy-terminal sequence of mouse Akt. Phospho-Akt1 (Ser473) (D7F10) XP® Rabbit mAb (Akt1 Specific) is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser473 of human Akt1 protein. Akt1 (C73H10) Rabbit mAb is produced by immunizing animals with a synthetic peptide surrounding Leu110 of human Akt1. Phospho-Akt2 (Ser474) (D3H2) Rabbit mAb (Akt2 Specific) is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser474 of human Akt2 protein. Akt2 (D6G4) Rabbit mAb is produced by immunizing animals with a synthetic peptide corresponding to residues of human Akt2.
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).
There are three Akt isoforms (Akt1, Akt2 and Akt3) in mammals (20). Akt activation requires phosphorylation by mTORC2 at Ser473 of Akt1, Ser474 of Akt2, and Ser472 of Akt3 (20).
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