Product Includes | Product # | Quantity | Color | Storage Temp |
---|---|---|---|---|
PRAS40 Mouse Detection mAb | 8075 | 1 ea |
|
+4C |
Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 1 ea |
|
+4C |
Detection Antibody Diluent | 13339 | 11 ml |
|
+4C |
HRP Diluent | 13515 | 11 ml |
|
+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 |
|
+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® Total PRAS40 Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of PRAS40. A PRAS40 rabbit antibody has been coated onto the microwells. After incubation with cell lysates, PRAS40 (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a PRAS40 mouse detection antibody is added to detect captured PRAS40 protein. Anti-mouse IgG, HRP-linked Antibody is then used to recognize the bound detection antibody. HRP substrate, TMB, is added to develop color. The magnitude of the absorbance for this developed color is proportional to the quantity of total PRAS40 protein.
Antibodies in kit are custom formulations specific to kit.
Specificity/Sensitivity
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
Limited Uses
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