Analysis Note

ControlA431cell lysate, or lysate of HeLa cells treated with heat shock
Included Positive Antigen Control: Catalog # 12-301, Non-stimulated A431 cell lysate. Add 2.5 µL of 2-mercaptoethanol/100 µL of lysate and boil for 5 minutes to reduce the preparation. Load 20 µg of reduced lysate per lane for minigels.

Application

Research CategorySignaling

Research Sub CategoryInsulin/Energy Signaling

Anti-AMPK alpha 1, Cat. No. 07-350 is rabbit polyclonal antibody that detects 5′-AMP-activated protein kinase catalytic subunit alpha-1 and is tested for use in Western Blotting and Immunohistochemistry.Western Blot Analysis: 2 µg/mL of this antibody detected AMPK α1 in RIPA lysates from non-stimulated A431 cells.Immunohistochemistry (Paraffin) Analysis: 1:250 dilution of this antibody detected AMPK α1 in human gallbladder tissue sections.

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

5′-AMP-activated protein kinase catalytic subunit alpha-1 (UniProt: Q13131; also known as EC:2.7.11.1, AMPK subunit alpha-1, Acetyl-CoA carboxylase kinase, ACACA kinase, Hydroxymethylglutaryl-CoA reductase kinase, HMGCR kinase, Tau-protein kinase PRKAA1) is encoded by the PRKAA1 (also known as AMPK1) gene (Gene ID: 5562) in human.AMPK is a heterotrimer composed of an alpha catalytic subunit (PRKAA1 or PRKAA2), a beta (PRKAB1 or PRKAB2) and a gamma non-catalytic subunits (PRKAG1, PRKAG2 or PRKAG3). It also contains an autoinhibitory sequence that shows some sequence similarity with the ubiquitin-associated domains and it represses kinase activity. AMPK is an energy sensor protein kinase that plays a key role in regulating cellular energy metabolism. In response to reduction of intracellular ATP levels, AMPK activates energy-producing pathways and inhibits energy-consuming processes, including inhibition of protein, carbohydrate and lipid biosynthesis, and reduces cell growth and proliferation. AMPK acts via direct phosphorylation of metabolic enzymes, and by longer-term effects via phosphorylation of transcription regulators. AMPK is shown to regulate insulin-signaling and glycolysis by phosphorylating IRS1, PFKFB2 and PFKFB3 and it stimulates glucose uptake in muscle by increasing the translocation of the glucose transporter GLUT4 to the plasma membrane. In the liver it acts as a key regulator of glucose homeostasis by phosphorylating CRTC2/TORC2, leading to CRTC2/TORC2 sequestration in the cytoplasm. AMPK is activated by phosphorylation on Thr183. Binding of AMP to non-catalytic gamma subunit results in allosteric activation, inducing phosphorylation on Thr183. AMP-binding to gamma subunit also sustains activity by preventing dephosphorylation of Thr183.

Immunogen

KLH-conjugated linear peptide corresponding to 17 amino acids from the C-terminal region of human AMPK catalytic subunit alpha-1.

Legal Information

UPSTATE is a registered trademark of Merck KGaA, Darmstadt, Germany

Linkage

Replaces: 04-323

Other Notes

Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.

Physical form

Protein A purified rabbit in buffer containing 0.1M Tris-Glycine, 0.15M NaCl, 0.05% Sodium Azide, pH 7.4.

Protein A chromatography

Format: Purified

Quality

Evaluated by immunoblot in RIPA lysates from non-stimulated A431 cells.

Western Blot Analysis:
0.5-2 µg/mL of this antibody detected AMPK α1 in RIPA lysates from non-stimulated A431 cells.

Specificity

This polyclonal antibody detects AMPK1 alpha subunit in human. It targets an epitope within 17 amino acids from the C-terminal half of human AMPK1.

Storage and Stability

Stable for 1 year at 2-8°C from date of receipt.

Target description

~63 kDa observed; 64.00 kDa calculated. Uncharacterized bands may be observed in some lysate(s).