Human/Mouse/Rat Total AMPK alpha 1 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 total AMPK alpha 1 in cell lysates. An immobilized capture antibody specific for AMPK alpha 1 binds both phosphorylated and unphosphorylated AMPK alpha 1. After washing away unbound material, a biotinylated detection antibody is used to detect both phosphorylated and unphosphorylated protein, utilizing a standard Streptavidin-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
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Quantification of AMPK alpha1 in human cell lysates. Lysates prepared from MCF-7 human breast cancer cells, MDA-MB-468 human breast cancer cells, and HeLa human cervical epithelial carcinoma cells were quantified with this DuoSet IC ELISA. The same lysates were also immunoblotted (inset) with anti-AMPK alpha1 polyclonal antibody (Catalog # AF3197). The DuoSet IC ELISA results correlate well with the relative amounts of AMPK alpha1 detected by Western blot.
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Quantification of AMPK alpha alpha1 in mouse and rat cell lysates. Lysates prepared from C2C12 mouse myoblast cells, DA3 mouse myeloma cells, and PC-12 rat adrenal pheochromocytoma cells were quantified with this DuoSet IC ELISA. The same lysates were also immunoblotted (inset) with anti-AMPK alpha1 monoclonal antibody (Catalog # MAB3197). The DuoSet IC ELISA results correlate well with the relative amounts of AMPK alpha1 detected by Western blot.
Product Datasheets
Preparation and Storage
Background: AMPK alpha 1
AMP-activated protein kinase (AMPK) is a heterotrimeric complex consisting of a catalytic a subunit and regulatory beta and gamma subunits. Each subunit exists as alternate isoforms (alpha 1, alpha 2, beta 1, beta 2, gamma 1, gamma 2, gamma 3), with all 12 combinations capable of forming complexes. The catalytic a subunit of AMPK is activated allosterically by AMP and by phosphorylation via the AMPK kinase LKB1. Active AMPK downregulates anabolic pathways such as fatty acid, triglyceride, and cholesterol synthesis, and up-regulates catabolic pathways such as glycolysis and fatty acid oxidation. AMPK's role in metabolic regulation has implicated this serine/threonine kinase as a therapeutic target in heart disease, obesity, and diabetes.
Citations for Human/Mouse/Rat Total AMPK alpha 1 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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Agomelatine improves streptozotocin-induced diabetic nephropathy through melatonin receptors/SIRT1 signaling pathway
Authors: NM Mahmoud, SM Elshazly, AA Hassan, E Soliman
International immunopharmacology, 2022-12-30;115(0):109646.
Species: Rat
Sample Types: Tissue Homogenates
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Metformin, an AMPK Activator, Inhibits Activation of FLSs but Promotes HAPLN1 Secretion
Authors: Y Chen, F Qiu, B Yu, Y Chen, F Zuo, X Zhu, KS Nandakumar, C Xiao
Mol Ther Methods Clin Dev, 2020-05-13;17(0):1202-1214.
Species: Human
Sample Types: Serum
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