REACTIVITY | SENSITIVITY | MW (kDa) | Isotype |
---|---|---|---|
H M R Mk Dm | Endogenous | Rabbit IgG |
Confocal immunofluorescent analysis of C2C12 cells, treated with LY294002 #9901 (50 μM, 2 hrs; left) or treated with insulin (100 ng/mL, 15 min; right), using Akt (pan) (C67E7) Rabbit mAb (Alexa Fluor® 647 Conjugate) (red pseudocolor) and Acetyl-Histone H3 (Lys9) (C5B11) Rabbit mAb (Alexa Fluor® 488 Conjugate) #9683 (green).
Learn more about how we get our imagesFlow cytometric analysis of Jurkat cells using Akt (pan) (C67E7) Rabbit mAb (Alexa Fluor® 647 Conjugate) (green) compared to Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate) #2985 (red).
Learn more about how we get our imagesNOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: Cells should be grown, treated, fixed and stained directly in multi-well plates, chamber slides or on coverslips.
Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.
NOTE: Formaldehyde is toxic, use only in a fume hood.
NOTE: All subsequent incubations should be carried out at room temperature unless otherwise noted in a humid light-tight box or covered dish/plate to prevent drying and fluorochrome fading.
posted November 2006
revised November 2013
Protocol Id: 182
NOTE: Prepare solutions with reverse osmosis deionized (RODI) or equivalent grade water.
NOTE: If using whole blood, lyse red blood cells and wash by centrifugation prior to fixation.
posted July 2009
revised June 2017
Protocol Id: 407
Application | Dilutions |
---|---|
Immunofluorescence (Immunocytochemistry) | 1:50 |
Flow Cytometry | 1:50 |
Supplied in PBS (pH 7.2), less than 0.1% sodium azide and 2 mg/ml BSA. Store at 4°C. Do not aliquot the antibody. Protect from light. Do not freeze.
Akt (pan) (C67E7) Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of total Akt protein. This antibody does not cross-react with other related proteins.
Human, Mouse, Rat, Monkey, D. melanogaster
Pig
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the carboxy terminus sequence of mouse Akt.
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. This antibody is expected to exhibit the same species cross-reactivity as the unconjugated Akt (pan) (C67E7) Rabbit mAb #4691.
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).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. The Alexa Fluor dye antibody conjugates in this product are sold under license from Life Technologies Corporation for research use only, except for use in combination with DNA microarrays. The Alexa Fluor® dyes (except for Alexa Fluor® 430 dye) are covered by pending and issued patents. Alexa Fluor® is a registered trademark of Molecular Probes, Inc. U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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Product # | Size | Price | Inventory |
---|---|---|---|
5186S | 100 µl (50 tests) |
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