Invitrogen™ Human AMPK alpha-1,2 (Phospho) [pT172] ELISA Kit

Description

ELISA

Kit is designed to detect and quantify the level of AMPK alpha-1,2 (Phospho) [pT172] in fresh or frozen human cell lysates. Cross-reactivity has been observed in mouse and rat cells. The assay recognizes both natural and recomb. human AMPK alpha-1,2 (Phospho) [pT172]. Principle of the method A monoclonal capture antibody specific for AMPK alpha-1,2 (Phospho) [pT172] has been coated onto the wells of 96-well plate. During the first incubation, standards of known content and unknown samples are pipetted into the wells and the antigen binds to the immobilized (capture) antibody. After washing, a rabbit antibody specific for the target protein is added to the wells and serves as a detection antibody by binding to the immobilized protein captured during the first incubation. After washing, a HRP labeled anti-rabbit IgG is added. This binds to the detection antibody to complete the four member sandwich. After a third incubation and washing to remove all the unbound enzyme, a substrate solution (TMB) is added, which is acted upon by the bound enzyme to produce color. The intensity of this colored product is directly proportional to the concentration of target protein present in the original specimen and the O.D. can be read on a standard microplate reader. Rigorous validation Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

The protein encoded by this gene is a catalytic subunit of the AMP-activated protein kinase (AMPK). AMPK is a heterotrimer consisting of an alpha catalytic subunit, and non-catalytic beta and gamma subunits. AMPK is an important energy-sensing enzyme that monitors cellular energy status. In response to cellular metabolic stresses, AMPK is activated, and thus phosphorylates and inactivates acetyl-CoA carboxylase (ACC) and beta-hydroxy beta-methylglutaryl-CoA reductase (HMGCR), key enzymes involved in regulating de novo biosynthesis of fatty acid and cholesterol. Studies of the mouse counterpart suggest that this catalytic subunit may control whole-body insulin sensitivity and is necessary for maintaining myocardial energy homeostasis during ischemia.

Specifications

• Accession Number: P54645,P54646,Q09137,Q13131,Q5EG47,Q8BRK8
• Assay Range: <1 U/mL
• Assay Sensitivity: <1 U/mL
• Concentration: 1.6 U/mL
• Conjugate: HRP
• Product Type: ELISA
• Sample Type: Cell Lysates
• Substrate Type: Serine/Threonine Kinases
• Target Species: Mouse, Rat
• For Use With (Equipment): Colorimetric Microplate Reader
• Gene ID (Entrez): 105787, 108079, 5562, 5563, 65248, 78975
• Gene Symbol: AMPK,AMPK2,AMPKa2,PRKAA,AMPK,AMPKa1
• Incubation Time: 4 hr.
• Interassay CV: 7.6%
• Intraassay CV: 4.8
• Kit Contents: Pre-coated 96 well plate, Standard, Standard Dilution Buffer, Detection Antibody, anti-rabbit-HRP, HRP Diluent, Wash Buffer, Chromogen, Stop Solution, Adhesive Plate Covers
• Label or Dye: HRP
• Marker Gene: AMPK
• Quantity: 96 Tests
• Regulatory Status: RUO
• Sample Volume: Cell Lysate 10 μL
• Storage Requirements: 2°C to 8°C
• Target: AMPK alpha-1,2
• Test Time: 1 hrs 20 mins
• Total Assay Time: 4 hrs