Summary
Product Features
Kit Content
Other Reagents Required
Data Examples
Product Datasheets
Preparation and Storage
Background
Phospho-Akt (S473) Pan Specific DuoSet IC ELISA Summary
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
Cell lysates (100 µL)
Assay Range
156.0 - 10,000 pg/mL
Sufficient Materials
Kits available for two, five, or fifteen 96-well plates*
Specificity
Please see the product datasheet
* 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, mouse, and rat Akt (S473) Pan Specific in cell lysates. An immobilized capture antibody specific for Akt binds both phosphorylated and unphosphorylated Akt. 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
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: Akt
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.
Long Name:
v-Akt Murine Thymoma Viral Oncogene Homolog
Alternate Names:
RAC-ALPHA; AKT serine/threonine kinase 1; Akt; PKB; PKB-ALPHA; PRKBA; RAC
Citations for Phospho-Akt (S473) Pan Specific 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.
16
Citations: Showing 1 - 10
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A small-molecule PI3K? activator for cardioprotection and neuroregeneration
Authors: Gong, GQ;Bilanges, B;Allsop, B;Masson, GR;Roberton, V;Askwith, T;Oxenford, S;Madsen, RR;Conduit, SE;Bellini, D;Fitzek, M;Collier, M;Najam, O;He, Z;Wahab, B;McLaughlin, SH;Chan, AWE;Feierberg, I;Madin, A;Morelli, D;Bhamra, A;Vinciauskaite, V;Anderson, KE;Surinova, S;Pinotsis, N;Lopez-Guadamillas, E;Wilcox, M;Hooper, A;Patel, C;Whitehead, MA;Bunney, TD;Stephens, LR;Hawkins, PT;Katan, M;Yellon, DM;Davidson, SM;Smith, DM;Phillips, JB;Angell, R;Williams, RL;Vanhaesebroeck, B;
Nature
Species: Human
Sample Types: Cell Lysates
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Identification and development of a subtype-selective allosteric AKT inhibitor suitable for clinical development
Authors: N Page, M Wappett, CR O'Dowd, M O'Rourke, G Gavory, L Zhang, JSS Rountree, L Jordan, O Barker, H Gibson, C Boyd, S Feutren-Bu, E McLean, G Trevitt, T Harrison
Scientific Reports, 2022-09-20;12(1):15715.
Species: Human
Sample Types: Cell Culture Supernates
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Decreased concentrations of intracellular signaling proteins in colon cancer patients with BRAF mutations
Authors: DA Olsen, CEB Thomsen, RF Andersen, JS Madsen, A Jakobsen, I Brandslund
Sci Rep, 2020-11-18;10(1):20113.
Species: Human
Sample Types: Tissue Homogenates
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Branched-chain ketoacid overload inhibits insulin action in the muscle
Authors: D Biswas, KT Dao, A Mercer, AM Cowie, L Duffley, Y El Hiani, PC Kienesberg, T Pulinilkun
J. Biol. Chem., 2020-09-02;0(0):.
Species: Mouse
Sample Types: Cell Culture Supernates
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Enhancing the regenerative effectiveness of growth factors by local inhibition of interleukin-1 receptor signaling
Authors: Z Julier, R Karami, B Nayer, YZ Lu, AJ Park, K Maruyama, GA Kuhn, R Müller, S Akira, MM Martino
Sci Adv, 2020-06-12;6(24):eaba7602.
Species: Mouse
Sample Types: Cell Culture Supernates
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Enhancing the regenerative effectiveness of growth factors by local inhibition of interleukin-1 receptor signaling
Authors: Z Julier, R Karami, B Nayer, YZ Lu, AJ Park, K Maruyama, GA Kuhn, R Müller, S Akira, MM Martino
Sci Adv, 2020;6(24):eaba7602.
Species: Mouse
Sample Types: Cell Culture Supernates
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A FUNCTIONAL ROLE OF GAS6/TAM IN NON-ALCOHOLIC STEATOHEPATITIS PROGRESSION IMPLICATES AXL AS THERAPEUTIC TARGET
Authors: A Tutusaus, E de Gregori, B Cucarull, H Cristóbal, C Aresté, I Graupera, M Coll, A Colell, G Gausdal, JB Lorens, P García de, A Morales, M Marí
Cell Mol Gastroenterol Hepatol, 2019-11-02;0(0):.
Species: Human
Sample Types: Cell Lysates
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Rosiglitazone accelerates wound healing by improving endothelial precursor cell function and angiogenesis in db/db mice
Authors: G Zhou, X Han, Z Wu, Q Shi, X Bao
PeerJ, 2019-10-17;7(0):e7815.
Species: Mouse
Sample Types: Cell Lysates
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Evidence that Graves' Ophthalmopathy Immunoglobulins Do Not Directly Activate IGF-1 Receptors
Authors: B Marcus-Sam, CC Krieger, A Boutin, G Kahaly, S Neumann, MC Gershengor
Thyroid, 2018-04-30;0(0):.
Species: Human
Sample Types: Cell Lysates
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Neurotensin stimulates sortilin and mTOR in human microglia inhibitable by methoxyluteolin, a potential therapeutic target for autism
Proc Natl Acad Sci USA, 2016-09-23;0(0):.
Species: Human
Sample Types: Cell Lysates
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The Escherichia coli subtilase cytotoxin A subunit specifically cleaves cell-surface GRP78 protein and abolishes COOH-terminal-dependent signaling.
Authors: Ray, Rupa, de Ridder, Gustaaf, Eu, Jerry P, Paton, Adrienne, Paton, James C, Pizzo, Salvator
J Biol Chem, 2012-07-31;287(39):32755-69.
Species: Human
Sample Types: Cell Lysates
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Protein secretion in human mammary epithelial cells following HER1 receptor activation: influence of HER2 and HER3 expression.
Authors: Zhang Y, Gonzalez RM, Zangar RC
BMC Cancer, 2011-02-14;11(0):69.
Species: Human
Sample Types: Cell Lysates
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Distinctive ERK and p38 signaling in remote and infarcted myocardium during post-MI remodeling in the mouse.
Authors: Yeh CC, Li H, Malhotra D, Turcato S, Nicholas S, Tu R, Zhu BQ, Cha J, Swigart PM, Myagmar BE, Baker AJ, Simpson PC, Mann MJ
J. Cell. Biochem., 2010-04-15;109(6):1185-91.
Species: Mouse
Sample Types: Tissue Homogenates
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Mechanism of the salutary effects of estrogen on kupffer cell phagocytic capacity following trauma-hemorrhage: pivotal role of Akt activation.
Authors: Hsieh CH, Nickel EA, Chen J, Schwacha MG, Choudhry MA, Bland KI, Chaudry IH
J. Immunol., 2009-04-01;182(7):4406-14.
Species: Mouse
Sample Types: Cell Lysates
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Citrullination of CXCL12 differentially reduces CXCR4 and CXCR7 binding with loss of inflammatory and anti-HIV-1 activity via CXCR4.
Authors: Struyf S, Noppen S, Loos T, Mortier A, Gouwy M, Verbeke H, Huskens D, Luangsay S, Parmentier M, Geboes K, Schols D, Van Damme J, Proost P
J. Immunol., 2009-01-01;182(1):666-74.
Species: Human
Sample Types: Cell Lysates
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Citrullination of CXCL10 and CXCL11 by peptidylarginine deiminase: a naturally occurring posttranslational modification of chemokines and new dimension of immunoregulation.
Authors: Loos T, Mortier A, Gouwy M, Ronsse I, Put W, Lenaerts JP, Van Damme J, Proost P
Blood, 2008-07-21;112(7):2648-56.
Species: Hamster
Sample Types: Cell Lysates
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Phospho-Akt (S473) Pan Specific DuoSet IC ELISA
By Jenna Nguyen
on 05/07/2019
Sample Tested: A7r5 rat thoracic aortic smooth muscle cell line
