Human/Mouse/Rat Total AMPK alpha 1 DuoSet IC ELISA

Name: Human/Mouse/Rat Total AMPK alpha 1 DuoSet IC ELISA
Brand: R&D Systems
SKU: DYC3197-2

R&D Systems, part of Bio-Techne | Catalog # DYC3197-2

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Product Summary
Product Specifications
Scientific Data
Kit Contents
Other Reagents Required
Preparation & Storage
Background
Product Documents
Specific Notices

Key Product Details

Assay Type

Solid Phase Sandwich ELISA

Assay Range

625-40000 pg/mL

Sample Type

Cell Lysates

Reactivity

Human, Mouse, Rat

Human/Mouse/Rat Total AMPK alpha 1 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

View all AMPK alpha 1 ELISA Kits »

Product Summary for Human/Mouse/Rat Total AMPK alpha 1 DuoSet IC ELISA

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 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

Please see the product datasheet

Label

HRP

Scientific Data Images for Human/Mouse/Rat Total AMPK alpha 1 DuoSet IC ELISA

Standard Curve

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.

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.

Kit Contents for Human/Mouse/Rat Total AMPK alpha 1 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

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

Alternate Names

PRKAA1

Entrez Gene IDs

5562 (Human)

Gene Symbol