Human/Mouse Phospho-Akt1 (S473) DuoSet IC ELISA Summary
* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated human and mouse Akt1 (S473) in cell lysates. An immobilized capture antibody specific for Akt1 binds both phosphorylated and unphosphorylated Akt1. After washing away unbound material, a biotinylated detection antibody is used to detect only phosphorylated receptor, utilizing a standard HRP format.
Product Features
- Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
- Development protocols are provided to guide further assay optimization
- Assay can be customized to your specific needs
- Available in 2, 5, and 15- (96-well) plate pack sizes
- Economical alternative to Western blot
Kit Content
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
- Streptavidin-HRP
Other Reagents Required
PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2O4, pH 7.2 - 7.4, 0.2 µm filtered
Wash Buffer: (Catalog # WA126), or equivalent
Lysis Buffer*
IC Diluent*
Blocking Buffer*
Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)
Stop Solution: 2 N H2SO4 (Catalog # DY994)
Microplates: From Costar EIA Plate (Costar Catalog # 2592) or R&D Systems (Catalog # DY990), or equivalent
Plate Sealers: ELISA Plate Sealers (Catalog # DY992), or equivalent
*For the Lysis Buffer, IC Diluent, and Blocking BUffer recommended for a specific DuoSet ELISA Development Kit, please see the product
Scientific Data
Product Datasheets
Preparation and Storage
Background: Akt1
Akt1, Akt2, and Akt3 make up a family of serine/threonine kinases, often activated downstream of growth factor receptors and PI 3-Kinase. Each Akt family member contains an N-terminal pleckstrin homology (PH) domain, a central kinase domain, and a C-terminal regulatory domain. They have putative roles in cell proliferation and survival, and Akt perturbations are thought to play an important role in tumorigenesis.
Citations for Human/Mouse Phospho-Akt1 (S473) DuoSet IC ELISA
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 6
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Microfluidic Single-Cell Proteomics Assay Chip: Lung Cancer Cell Line Case Study
Authors: Y Jung, M Son, YR Nam, J Choi, JR Heath, S Yang
Micromachines, 2021-09-23;12(10):.
Species: Human
Sample Types: Cell Lysates
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Cetuximab sensitivity of head and neck squamous cell carcinoma xenografts is associated with treatment-induced reduction of EGFR, pEGFR, and pSrc
Authors: Adam Jedlinski
J. Oral Pathol. Med, 2017-01-28;0(0):.
Species: Human
Sample Types: Cell Lysates
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Natural nitration of CXCL12 reduces its signaling capacity and chemotactic activity in vitro and abrogates intra-articular lymphocyte recruitment in vivo
Authors: R Janssens, A Mortier, D Boff, V Vanheule, M Gouwy, C Franck, O Larsen, MM Rosenkilde, J Van Damme, FA Amaral, MM Teixeira, S Struyf, P Proost
Oncotarget, 2016-09-20;7(38):62439-62459.
Species: Mouse
Sample Types: Cell Culture Supernates
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Stand-Sit Microchip for High-Throughput, Multiplexed Analysis of Single Cancer Cells
Sci Rep, 2016-09-01;6(0):32505.
Applications: ChIP -
Real-time monitoring of Cisplatin-induced cell death.
Authors: Alborzinia H, Can S, Holenya P, Scholl C, Lederer E, Kitanovic I, Wolfl S
PLoS ONE, 2011-05-16;6(5):e19714.
Species: Human
Sample Types: Cell Lysates
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Proteolytic processing of CXCL11 by CD13/aminopeptidase N impairs CXCR3 and CXCR7 binding and signaling and reduces lymphocyte and endothelial cell migration.
Authors: Proost P, Mortier A, Loos T, Vandercappellen J, Gouwy M, Ronsse I, Schutyser E, Put W, Parmentier M, Struyf S, Van Damme J
Blood, 2007-03-15;110(1):37-44.
Species: Hamster
Sample Types: Cell Lysates
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