Human Phospho-AMPK alpha 1 (T183) DuoSet IC ELISA

Catalog # Availability Size / Price Qty
DYC3528-5
DYC3528-2
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Human Phospho-AMPK alpha 1 (T183) 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)
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 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 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

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

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.

Long Name:
AMP-activated Protein Kinase alpha 1
Entrez Gene IDs:
5562 (Human)
Alternate Names:
AMP -activate kinase alpha 1 subunit; AMP-activated protein kinase, catalytic, alpha-1,5'-AMP-activated protein kinase catalytic subunit alpha-1; AMPK alpha 1; AMPK alpha 1,5'-AMP-activated protein kinase, catalytic alpha-1 chain; AMPK subunit alpha-1; AMPK; AMPK1; AMPKa1; EC 2.7.11; EC 2.7.11.1; MGC33776; MGC57364; PRKAA1; protein kinase, AMP-activated, alpha 1 catalytic subunit

FAQs

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