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Invitrogen
Multispecies AKT Pathway (AKT/GSK/p70 S6K) Activation Profile InstantOne™ ELISA Kit
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Multispecies AKT Pathway (AKT/GSK/p70 S6K) Activation Profile InstantOne™ ELISA Kit
Representative data for AKT Pathway (AKT/GSK/p70 S6K) Activation Profile Multispecies InstantOne™ ELISA. Representative data for p38 MAPK (Total) Multispecies InstantOne™ ELISA. MCF7 cells were treated with either the PI3 Kinase inhibitor Wortmannin (10 µM) (-) or insulin (10 ng/mL for 10 min.) (+). Cell lysates were prepared as described in the InstantOne™ ELISA User Manual. Using the AKT Pathway Activa... View More
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Product Specifications
Sample type/volume
Cell lysate
20 µL
Hands-on time
1 hr 20 min
Time-to-result
1 hr 10 min
Homogenous (no wash)
No
Instrument
Absorbance Microplate Reader
Product size
96 Tests
Contents
Capture Antibody
Detection Antibody Reagent
InstantOne™ Assay Plate
Cell Lysis Buffer
Enhancer Solution
Wash Buffer (10X)
Substrate Reagent (TMB)
Stop Solution
Positive Control Cell Lysate (lyophilized)
Plate seal
Detection Antibody Reagent
InstantOne™ Assay Plate
Cell Lysis Buffer
Enhancer Solution
Wash Buffer (10X)
Substrate Reagent (TMB)
Stop Solution
Positive Control Cell Lysate (lyophilized)
Plate seal
Shipping conditions
Wet ice
Storage
2-8°C
Protein name
AKT Pan/GSK3 alpha/beta/p70 S6 Kinase
Target site-specificity
Detector antibody conjugate
Biotin
Label or dye
HRP
About This Kit
The InstantOne™ ELISA is specifically engineered for accurate measurement of phosphorylated AKT 1/2/3 (Ser473) (human, mouse, hamster, rat), phosphorylated GSK3beta (Ser9) (human, mouse, rat) and phosphorylated p70S6K (Thr389) (human, mouse) in cell lysates. The InstantOne™ ELISA kit allows for fast analysis of samples in approximately one hour. All reagents used in a traditional sandwich ELISA are added in solution to a plate followed by a wash step and detection with the TMB colorimetric substrate.
Target information
AKT, which exists as multiple isoforms, is one of the principle kinases activated by phosphoinositide 3-kinase (PI3K). Activation of PI3K results in the generation of phosphatidylinositol (3,4,5)-triphosphate (PIP3). These lipid second messengers bind to the pleckstrin homology domain of AKT to promote its translocation to the plasma membrane for activation via phosphorylation at Thr308 and Ser473 by PDK1 and the mTOR TORC2 complex, respectively. Phosphorylation at both these sites is required for full activation of AKT Ser/Thr kinase activity. AKT phosphorylates over 50 known substrates, including GSK3b, p70S6K, AS160, PRAS40, TSC 1, TCS 2, Raf-1, Bad, the FOXO family of transcription factors, and PFK2. Due to its established critical role in numerous biological processes, AKT is a frequent target for oncology-related drug development. Glycogen Synthase Kinase 3 (GSK3) is a Ser/Thr kinase that exists as two isoforms, GSK3a and GSK3β. Unlike other kinases, phosphorylation at Ser9 and Ser21 for GSK3alpha and GSK3beta, respectively, leads to inactivation of this enzyme. GSK3 has been implicated in several diseases, including cancer and diabetes, through its phosphorylation of glycogen synthase. GSK3 has also been associated with neurodegenerative diseases such as Alzheimer's disease due to its phosphorylation of the neuronal Tau protein. p70 S6 Kinase (p70S6K) is a member of the ribosomal S6 kinase family of Ser/Thr kinases. P70 S6K activity is controlled by multiple signaling pathways, including the MAPK, phosphoinositide-3 kinase (PI3K), and mTOR pathways. This regulation results in the phosphorylation of multiple sites of p70 S6K, including Thr389 within the linker domain, which is required for full activation. Activated p70 S6K phosphorylates several residues on the S6 ribosomal protein, thereby leading to increased protein synthesis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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If an Invitrogen™ immunoassay doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*
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