AKT1 STAR ELISA Kit
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EMD Millipore
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Product
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AKT1 STAR ELISA Kit
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Company
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EMD Millipore
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Price
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More Information
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View Company Product Page
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Accession Number
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NM_001014431.1,NM_001014432.1
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Applications
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ELISA
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Detection
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Chromogenic;Chromogenic
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Form
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Routinely evaluated by ELISA.
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Reactivity
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Human;Mouse;Rat
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Source/Expression System
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Immunology or serology test kits or supplies
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Target
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AKT1
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Catalog Number
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17-455
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Method
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ELISA
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Original Item Name
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AKT1 STAR ELISA Kit
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Quantity
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96 assays
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Sensitivity
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Sensitivity: 0.3 ng/mL. Range of Detection: 0.3 to 20 ng/mL
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Target/Molecule Descriptor
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STAR
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Description
AKT1 STAR ELISA Kit The UPSTATE colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The AKT1 plate is coated with a specific mouse monoclonal anti-AKT1 capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standard included in the kit are incubated in the microwells allowing AKT1 antigen to be captured in the plate wells. The plate is then washed to remove any non-bound unspecific material. The wells are then incubated with a specific rabbit anti-AKT1 antibody to detect the captured AKT1 on the plate well. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. This allows for a sensitive enzymatic detection of the sample. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader. The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to develop a standard curve. II. Akt BACKGROUND Akt (Protein Kinase B), a Ser/Thr kinase, is a major known effecter of the PI3 Kinase pathway and is involved in multiple signaling pathways that relate to many biological processes including glucose metabolism, cell survival/apoptosis, cell cycle control, angiogenesis, differentiation, and cell growth and proliferation. Akt is activated by ligand-stimulated growth factor receptor signaling that activates the Phosphatidylinositol 3-kinase (PI3 Kinase, PI3K) dependent manner. PKB is one of the most frequently hyperactivated protein kinases in human cancers. In mammals three isoforms of Akt (Akt1/PKBα, Akt2/PKBβ, and Akt3/PKBγ) exists. They exhibit a high degree of homology, but differ slightly in the localization of their regulatory phosphorylation sites. Akt1 is the predominant isoform that is in most tissues and is thought to have a dominant role in growth, survival, embryonic development, and post-natal survival. Additionally, Akt1/PKBα is required for adipocyte differentiation, where as Akt2/PKBβ and Akt3/PKBγ are not. Akt2 is strongly correlated with the regulation of glucose homeostasis and is the predominant PKB isoform expressed in insulin-responsive tissues where defective Akt2 results in impaired insulin-stimulated glucose uptake in muscle and adipocytes. Akt3 is abundant in brain tissue. Each Akt isoform is composed of three functionally distinct regions: an N-terminal Pleckstrin Homology (PH) domain that provides a lipid-binding module to direct Akt to PIP3 at the cell membrane as a result of PI3 Kinase (PI3K) activity that is necessary for its activation, a central catalytic domain, and a C-terminal hydrophobic motif. The activation and regulation of AKT is dependent on a dual regulatory mechanism that requires both its translocation to the plasma membrane and dual phosphorylation on Thr308 and Ser473 by PDK1 and the TORC2 complex, respectively. This is accomplished by the generation and build-up of PIP3 by PI3K in conjunction with reduced PTEN function that results in the activation of PDK1 (3-phosphoinositide-dependent protein kinase-1) and the recruitment of AKT to the plasma membrane by direct interaction with its PH domain. The activated PDK1 then in turn phosphorylates Akt on Thr308 in its activation loop. This phosphorylation is necessary and sufficient for AKT activation; however maximal activation requires the additional phosphorylation at Ser473. Another kinase complex, recently identified as TORC2, which is composed of the mTOR, Rictor, Gï¢L, Sin1, and Protor1 and 2 (previously referred to as the unidentified kinase PDK2), phosphorylates AKT on Ser473
Specifications/Features
1. Capture Plate pre-coated with anti-AKT1 antibody: (Part No. 17-455A) One pre-coated 96-stripwell immunoplate sealed in a foil pouch.;2. Anti-AKT1 detection antibody: (Part No. 17-455B) One bottle (11 mL) of anti-AKT1 detection antibody containing sodium azide, ready to use.;3. ELISA Diluent: (Part No. 17-455C) One bottle (25 mL) of ELISA Diluent containing sodium azide, ready to use.;4. 25X ELISA Wash Buffer: (Part No. 17-455D) One bottle (50 mL) of 25X ELISA Wash Buffer;5. Anti-Rabbit IgG HRP conjugate: (Part No. 17-455E) One vial (125 &micor;L) of 100X anti-rabbit HRP conjugate containing thimerosol;6. HRP Diluent: (Part No. 17-455F) One bottle (25 mL) of HRP Diluent containing thimerosol;7. TMB Solution: (Part No. 17-455G) One bottle (25 mL) of stabilized tetramethylbenzidine (TMB), ready to use;8. Stop Solution: (Part No. 17-455H) One bottle (25 mL) of stop solution, ready to use.;9. AKT1 Standard: (Part No. 17-455I) Four vials of AKT1 standard, lyophilized;10. Plate Covers: Two plate covers.