Technical guide & protocol Tech Support

Product Specifications

Analytical sensitivity

<1 U/mL

Assay range

< 1 U/mL

Sample type/volume

Cell Lysate
10 µL

Hands-on time

1 hr 20 min

Time-to-result

4 hr

Homogenous (no wash)

No

Interassay CV

7.6%

Intraassay CV

4.8%

Instrument

Colorimetric Microplate Reader

Product size

96 Tests

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

Shipping conditions

Wet ice

Storage

2-8°C

Protein name

AMPK alpha-1,2

Target site-specificity

Phosphorylated Thr172

Protein subtype

Serine/Threonine Kinases

Protein family

Kinases & Inhibitors

Species (tested)

Human, Mouse, Rat

Assay kit format

Sandwich ELISA Kit

Detector antibody conjugate

HRP

Label or dye

HRP

About This Kit

The Human AMPK alpha-1,2 (Phospho) [pT172] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) 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 recombinant 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 the 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 horseradish peroxidase 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 optical density 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.

Target information

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.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Bioinformatics

Gene aliases : 2310008I11Rik, A830082D05, AI194361, AI450832, AL024255, AMPK, AMPK1, AMPK2, AMPKa1, AMPKa2, AMPKalpha1, AMPKalpha2, C130083N04Rik, PRKAA, PRKAA1, PRKAA2

Gene ID : (Human) 5562, (Human) 5563, (Rat) 65248, (Mouse) 105787, (Rat) 78975, (Mouse) 108079

Gene symbol : PRKAA1, PRKAA2, Prkaa1, Prkaa1, Prkaa2, Prkaa2

Protein Aliases : 5'-AMP-activated protein kinase catalytic subunit alpha-1, 5'-AMP-activated protein kinase catalytic subunit alpha-2, 5'-AMP-activated protein kinase, catalytic alpha-1 chain, catalytic alpha-2 chain, ACACA kinase, Acetyl-CoA carboxylase kinase, AMP -activate kinase alpha 1 subunit, AMP-activated protein kinase, AMP-activated protein kinase alpha-2 subunit variant 2, AMP-activated protein kinase alpha-2 subunit variant 3, AMP-activated protein kinase alpha-2 variant B, alpha 1 catalytic subunit, catalytic, alpha-1, AMPK alpha 1, AMPK alpha 2, AMPK alpha-1 chain, AMPK alpha-2 chain, AMPK subunit alpha-1, AMPK subunit alpha-2, AMPK-alpha-2 chain, HMGCR kinase, Hydroxymethylglutaryl-CoA reductase kinase, prkaa2, protein kinase, AMP-activated, alpha 2 catalytic subunit, Tau-protein kinase PRKAA1

UniProt ID (Human) Q13131, (Human) P54646, (Rat) P54645, (Mouse) Q5EG47, (Rat) Q09137, (Mouse) Q8BRK8

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