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PRAS40 STAR ELISA Kit
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PRODUCT FAMILY INFORMATION
STAR ELISA Kits
Upstate STAR (Signal Transduction Assay Reaction) ELISA kits are a fast, sensitive method to detect the relative amounts of total and activated targets through the use of phospho-specific antibodies.
Upstate STAR (Signal Transduction Assay Reaction) ELISA kits are a fast, sensitive method to detect the relative amounts of total and activated targets through the use of phospho-specific antibodies.
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Description:
PRAS40 STAR ELISA Kit
Trade Name:
Upstate (Millipore)
Qty/Pk:
96 assays
Background Information:
I. TEST PRINCIPLE
The UPSTATE® colorimetric STAR (Signal Transduction Assay Reaction) ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to detect specific levels of signaling targets in whole cell extracts. The PRAS40 plate is coated with a specific mouse monoclonal PRAS40 capture antibody on the microwells of the 96-well clear plate. Sample lysate or the standard included in the kit are incubated in the microwells allowing PRAS40 antigen to be captured in the plate wells. The plate is then washed to remove any unbound non-specific material. The wells are then incubated with a specific rabbit anti-phospho-PRAS40 (Thr246) antibody to detect the captured PRAS40 on the plate well that is phosphorylated on Thr246. The unbound detection antibody is washed away followed by incubation with an HRP-conjugated anti-rabbit antibody. After the addition of TMB substrate and stop solution the absorbance is measured at 450 nm using a plate reader. This allows for a sensitive enzymatic detection of the sample.
The entire assay takes less than 5 hours to complete with minimal hands-on time. Many of the reagents are supplied in ready-to use formulations for ease of use. The kit also includes a standard that is run as both a positive control and to generate a standard curve for phosphorylated PRAS40 (Thr246) measurement.
II. PRAS40 BACKGROUND
PRAS40 (Proline Rich Akt Substrate, 40kDa), also known as AKT1S1, is a ubiquitously express protein that is phosphorylated on residue Thr246 via the PI3K/Akt pathway. It was originally discovered as an Akt substrate as it has the putative Akt phosphorylation motif of RxRxxpS/pT2. This phosphorylation allows it to bind 14-3-3 and Raptor of the mTOR complex mTORC1. PRAS40 is known to bind to and regulate mTORC1 (mTOR, Raptor, mLST8) kinases activity that is activated by insulin downstream of PI3K and Akt and subsequently phosphorylates p70S6K and 4EBP1. PRAS40 is known to have a putative TOS motif (FVMDE) that allows it bind to Raptor of mTORC1 in the absence of insulin. It is thought that through it binding to the Raptor, PRAS40 inhibits the kinase activity of mTORC1 by preventing to bind to its substrates such as p70S6K and 4EBP1.
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Applications:
The PRAS40 colorimetric STAR ELISA kit is a solid phase sandwich enzyme linked immunosorbent assay that provides a fast, sensitive method to quantify specific levels of signaling targets in denatured cell extracts.
Key Applications:
ELISA
Species Reactivity:
Human
Usage Statement:
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.
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Entrez Gene Summary:
AKT1S1 is a proline-rich substrate of AKT (MIM 164730) that binds 14-3-3 protein (see YWHAH, MIM 113508) when phosphorylated (Kovacina et al., 2003 [PubMed 12524439]).[supplied by OMIM]
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