Documents

CS0160

Sigma-Aldrich

Akt/PKB ELISA

sufficient for 96 tests

Pricing and availability is not currently available.

usage

sufficient for 96 tests

shipped in

wet ice

storage temp.

2-8°C

Gene Information

human ... AKT1(207)

Looking for similar products? Visit Product Comparison Guide

General description

The assay is a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA). A monoclonal capture antibody specific for the Akt/PKB, regardless of phosphorylation state, has been coated onto the multiwell strips provided with the kit. Standard dilutions and samples are incubated for 2 hours at RT. Akt/PKB antigen binds to the capture antibody. After a wash, a detection antibody specific for the non-phosphorylated or phosphorylated protein is incubated for 1 hr at RT, which results in binding to the immobilized Akt/PKB protein. An anti-rabbit IgG-HRP completes the four-member sandwich. The reaction is visualized by tetramethylbenzidine (TMB) substrate, followed by the stop solution. The intensity of the yellow color, measured in a multiwell plate reader at 450 nm, is directly proportional to the concentration of Akt/PKB in the original sample. The unknown concentrations are calculated from the standard curve run with each assay.

Application

Akt/PKB ELISAs are designed for in vitro quantitative determination of human, mouse and rat Akt/PKB protein in cell lysates. Akt, also known as protein kinase Bα (PKBα) or RAC-PKa, is one of the downstream targets of PI3 Kinase (PI3K). Akt consist of three highly conserved isoforms designated in humans as Akt1, Akt2, and Akt3. Activated Akt acts as a key mediator of signals for cell survival, proliferation, angiogenesis, and a number of metabolic insulin effects. Because of its growth-promoting effects, Akt plays a central role in tumorigenesis. A number of oncogenes and tumor suppressor genes act upstream of Akt to influence cancer progression.

Biochem/physiol Actions

Akt/PKB ELISA is designed to detect and quantify the levels of human, mouse and rat Akt/PKB protein, independent of its phosphorylation state. Akt is one of the downstream targets of PI-3 Kinase (PI3-K). Akt consist of three highly conserved isoforms, designated in humans as Akt1, Akt2, and Akt3. Each isoform consists of an N-terminal pleckstrin homology (PH) domain, which mediates lipid-protein or protein-protein interactions, and a C-terminal kinase catalytic domain. All isoforms respond in a similar fashion to various stimuli. Akt can be activated by growth factors and physiologic stimuli in a PI3-K-dependent manner. Activation of Akt kinase involves both membrane translocation and phosphorylation.

Features and Benefits

Akt/PKB ELISAs are a fast (total time 4 hours), sensitive (<0.1 ng/ml for non-phosphorylated and <0.8 units/mL for phosphorylated Akt/PKB) and specific (measure total or phosphorylated Akt/PKB with no crossreactivity with p38 MAPK, p42, JNK1 and others) alternative to immunoblotting or bioassays. The kit contains precoated plates and incubations are run at room temperature. No proprietary instrumentation is required.

Analysis Note

The sensitivity is <0.1 ng/mL, which is ~2x more sensitive than immunoblotting. The assay is specific for total Akt/PKB and does not crossreact with p38 MAPK, p42 Erk1, p42 Erk2, JNK 1, human and rat insulin receptor, or human EGFR.

Safety Statement

24/25

RIDADR

UN 3316 9/PG 3

Certificate of Analysis
Certificate of Origin
André Bortolini Silveira et al.
Oncotarget, 6(15), 13105-13118 (2015-04-15)
The PI3K pathway is frequently hyperactivated in primary T-cell acute lymphoblastic leukemia (T-ALL) cells. Activation of the PI3K pathway has been suggested as one mechanism of glucocorticoid resistance in T-ALL, and patients harboring mutations in the PI3K negative regulator PTEN...
Dexin Kong et al.
Investigational new drugs, 32(6), 1134-1143 (2014-08-26)
As accumulating evidences suggest close involvement of phosphatidylinositol 3-kinase (PI3K) in cancer, novel PI3K inhibitors such as ZSTK474, GDC-0941, NVP-BEZ235 and BKM-120 have been developed for cancer therapy. A high frequency of hotspot mutations known as E542K, E545K and H1047R...
Abolfazl Avan et al.
PloS one, 9(9), e108057-e108057 (2014-09-23)
Pancreatic ductal adenocarcinoma (PDAC) patients have the highest risk of developing cachexia, which is a direct cause of reduced quality of life and shorter survival. Novel biomarkers to identify patients at risk of cachexia are needed and might have a...
Andrew M Cheng et al.
American journal of physiology. Endocrinology and metabolism, 307(7), E571-E579 (2014-08-15)
Among the pleotropic effects of endothelial nitric oxide (NO) is protection against vascular inflammation during high-fat diet (HFD) feeding. The current work investigated the role of the enzyme vasodilatory-stimulated phosphoprotein (VASP) as a downstream mediator of the anti-inflammatory effect of...
Matias Juanes et al.
Clinical endocrinology, 82(5), 704-711 (2014-07-22)
IGF1R gene mutations have been associated with varying degrees of intrauterine and postnatal growth retardation, and microcephaly. To identify and characterize IGF1R gene variations in a cohort of 28 Argentinean children suspected of having IGF-1 insensitivity, who were selected on...

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon

MilliporeSigma

Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.