PathScan® Phospho-PRAS40 (Thr246) Sandwich ELISA Kit (#7327) Datasheet with Images Cell Signaling Technology

For Research Use Only. Not for Use in Diagnostic Procedures.

Product Includes	Product #	Quantity	Color	Storage Temp
PRAS40 Rabbit mAb Coated Microwells	82334	96 tests		+4C
Phospho-PRAS40 (Thr246) Rabbit Detection mAb (Biotinylated)	8196	1 ea	Green (Lyophilized)	+4C
HRP-Linked Streptavidin (ELISA Formulated)	11805	1 ea	Red (Lyophilized)	+4C
Detection Antibody Diluent	13339	11 ml	Green	+4C
HRP Diluent	13515	11 ml	Red	+4C
TMB Substrate	7004	11 ml		+4C
STOP Solution	7002	11 ml		+4C
Sealing Tape	54503	2 ea		+4C
ELISA Wash Buffer (20X)	9801	25 ml		+4C
ELISA Sample Diluent	11083	25 ml	Blue	+4C
Cell Lysis Buffer (10X)	9803	15 ml		-20C

*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.

Description

The PathScan^® Phospho-PRAS40 (Thr246) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PRAS40 when phosphorylated at Thr246. A PRAS40 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, PRAS40 protein (phosphorylated and nonphospho) is captured by the coated antibody. Following extensive washing, a biotinylated phospho-PRAS40 (Thr246) rabbit detection antibody is added to detect the captured phospho-PRAS40 (Thr246) protein. HRP-linked streptavidin is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for the developed color is proportional to the quantity of PRAS40 phosphorylated at Thr246.

*Antibodies in this kit are custom formulations specific to kit.

Specificity/Sensitivity

PathScan^® Phospho-PRAS40 (Thr246) Sandwich ELISA Kit #7327 detects endogenous levels of PRAS40 protein when phosphorylated at Thr246 as shown in Figure 1. Kit sensitivity is shown in Figure 2. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

Background

Many growth factors and hormones induce the phosphoinositide 3-kinase signaling pathway, which results in the activation of downstream effector proteins such as the serine/threonine kinase Akt (1,2). One known Akt substrate is a 40 kDa, proline-rich protein (PRAS40) that binds to 14-3-3 proteins (2). PRAS40 also binds mTOR to transduce Akt signals to the mTOR complex. Inhibition of mTOR signaling stimulates PRAS40 binding to mTOR, which in turn inhibits mTOR activity (3). PRAS40 interacts with raptor in mTOR complex 1 (mTORC1) in insulin-deprived cells and inhibits the activation of the mTORC1 pathway mediated by the cell cycle protein Rheb. Phosphorylation of PRAS40 by Akt at Thr246 relieves PRAS40 inhibition of mTORC1 (4). mTORC1 in turn phosphorylates PRAS40 at Ser183 (5).

Background References

Cross-Reactivity Key

H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected

Trademarks and Patents

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.

PathScan is a registered trademark of Cell Signaling Technology, Inc.

U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.

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