Human Phospho-AMPK alpha 1 (T183) 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.
- 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
- Capture Antibody
- Conjugated Detection Antibody
- Calibrated Immunoassay Standard or Control
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
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
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.
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