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.
Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA
R&D Systems | Catalog # DYC3528-2
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Key Product Details
Assay Type
Solid Phase Sandwich ELISA
Assay Range
62.5-4000 pg/mL
Sample Type
Cell Lysates
Reactivity
Human
Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA 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
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Product Summary for Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA
This DuoSet IC ELISA contains the basic components required for the development of sandwich ELISAs to measure phosphorylated human AMPK alpha 1 (T183) 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 only phosphorylated receptor, utilizing a standard HRP format.
Product Specifications
Assay Format
96-well strip plate (sold separately)
Sample Volume Required
Cell lysates (100 µL)
Detection Method
Colorimetric ELISA - 450nm (TMB)
Conjugate
Biotin
Specificity
Label
HRP
Kit Contents for Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA
- 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
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: AMPK alpha 1
Long Name
AMP-activated Protein Kinase alpha 1
Alternate Names
PRKAA1
Entrez Gene IDs
5562 (Human)
Gene Symbol
PRKAA1
Additional AMPK alpha 1 Products
Product Documents for Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA
Product Specific Notices for Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA
For research use only
Related Research Areas
Citations for Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- ELISA Sample Preparation & Collection Guide
- ELISA Troubleshooting Guide
- How to Run an R&D Systems DuoSet ELISA
- How to Run an R&D Systems Quantikine ELISA
- How to Run an R&D Systems Quantikine™ QuicKit™ ELISA
- Quantikine HS ELISA Kit Assay Principle, Alkaline Phosphatase
- Quantikine HS ELISA Kit Principle, Streptavidin-HRP Polymer
- Sandwich ELISA (Colorimetric) – Biotin/Streptavidin Detection Protocol
- Sandwich ELISA (Colorimetric) – Direct Detection Protocol
- Troubleshooting Guide: ELISA
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
