Product Pathways - PathScan ELISA
PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Antibody Pair #7854
| Kit Includes | Volume | Cap Color |
|---|---|---|
| 4E-BP1 (Thr37/46) Rabbit Capture Antibody (100X) | 0.4 milliliters | Pink |
| 4E-BP1 Mouse Detection Antibody (100X) | 0.4 milliliters | Blue |
| Anti-Mouse IgG, HRP-Linked Antibody (1000X) | 0.04 milliliters | Yellow |
Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.
Species Cross-Reactivity
H M R Mk
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey
Description
CST's PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Kit #7216. Capture and Detection antibodies (100X stocks) and HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Phospho-4E-BP1 (Thr37/Thr46) Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a 4E-BP1 Detection Antibody and anti-Mouse IgG, HRP conjugated antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of Phospho-4E-BP1 (Thr37/Thr46) protein.*Antibodies in this kit are custom formulations specific to the kit.
Sandwich ELISA
The relationship between the protein concentration of lysate from amino acid (AA)/untreated and AA/insulin-treated HEK- 293T cells and the absorbance at 450 nM using PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Antibody Pair #7854 is shown. HEK-293T cells were serum-starved overnight and deprived of amino acids for 1 hour. The amino acids were replenished for 1 hour. Cells were either untreated or stimulated with 100 nM insulin for 30 minutes at 37°C.
Background
Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the eIF4E translation initiation factor. Hyperphosphorylation of 4E-BP1 disrupts this interaction and results in activation of cap-dependent translation (1). Both the PI3 kinase/Akt pathway and FRAP/mTOR kinase regulate 4E-BP1 activity (2,3). Multiple 4E-BP1 residues are phosphorylated in vivo (4). While phosphorylation by FRAP/mTOR on Thr37 and Thr46 does not prevent the binding of 4E-BP1 to eIF4E, it is thought to prime 4E-BP1 for subsequent phosphorylation at Ser65 and Thr70 (5).
- Pause, A. et al. (1994) Nature 371, 762-767.
- Brunn, G.J. et al. (1997) Science 277, 99-101.
- Gingras, A.C. et al. (1998) Genes Dev. 12, 502-513.
- Fadden, P. et al. (1997) J. Biol. Chem. 272, 10240-10247.
- Gingras, A.C. et al. (1999) Genes Dev. 13, 1422-1437.
Application References
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
Companion Products
- 9808 Phosphate Buffered Saline (PBS-20X)
- 9809 Phosphate Buffered Saline with Tween 20 (PBST-20X)
- 9998 BSA
- 7004 TMB Substrate
- 7002 STOP Solution
- 9803 Cell Lysis Buffer (10X)
- 7076 Anti-mouse IgG, HRP-linked Antibody
- 7216 PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Kit
- 9644 4E-BP1 (53H11) Rabbit mAb
- 9452 4E-BP1 Antibody
- 2855 Phospho-4E-BP1 (Thr37/46) (236B4) Rabbit mAb
- 4923 Nonphospho-4E-BP1 (Thr46) (87D12) Rabbit mAb
- 9459 Phospho-4E-BP1 (Thr37/46) Antibody
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
