Technical guide & protocol Tech Support

Product Specifications

Analytical sensitivity

<0.5 U/mL

Assay range

1.6-100 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

5.9%

Intraassay CV

4.7%

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

PRAS40

Target site-specificity

Phosphorylated Thr246

Protein subtype

Adaptor Proteins

Protein family

Kinases & Inhibitors

Species (tested)

Human

Assay kit format

Sandwich ELISA Kit

Detector antibody conjugate

HRP

Label or dye

HRP

About This Kit

The Human PRAS40 (Phospho) [pT246] ELISA Kit is a solid-phase sandwich Enzyme-Linked Immunosorbent Assay (ELISA) designed to detect and quantify the level of PRAS40 (Phospho) [pT246] in fresh or frozen human cell lysates. The assay recognizes both natural and recombinant human PRAS40 (Phospho) [pT246].

Principle of the method

A monoclonal capture antibody specific for PRAS40 (Phospho) [pT246] 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 Akt signaling pathway contributes to the regulation of apoptosis after a variety of cell death signals. AKT1S1, also known as PRAS40, is a proline-rich substrate of the kinase AKT1 and is thought to play a role in neuroprotection mediated by nerve growth factor (NGF) after transient focal cerebral ischemia. AKT1S1 is also a substrate and potential regulator of mammalian target of rapamycin (mTOR), a serine/threonine kinase that regulates cell growth and cell cycle, and a negative regulator of autophagy. Treatment with the insulin-like growth factor-1 (IGF1) can induce the phosphorylation of AKT1S1 via the PI3K/AKT signaling pathway in PC12 cells.

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

Bioinformatics

Gene aliases : AKT1S1, Lobe, PRAS40

Gene ID : (Human) 84335

Gene symbol : AKT1S1

Protein Aliases : 40 kDa proline-rich AKT substrate, AKT1 substrate 1 (proline rich), AKT1 substrate 1 (proline-rich), AKT1S1, Proline-rich AKT1 substrate 1

UniProt ID (Human) Q96B36

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