Product Pathways - PathScan ELISA
PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Antibody Pair #7854
| Kit Includes | Volume | Cap Color |
|---|---|---|
| 4E-BP1 (T37/46) Capture Ab (100X) | 0.4 ml | Pink |
| 4E-BP1 Detection Ab (100X) | 0.4 ml | Blue |
| Anti-mouse IgG, HRP-linked Antibody | 0.04 ml | Yellow |
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Species Cross-Reactivity
H M R Mk
Reactivity Key:
H=Human
M=Mouse
R=Rat
Mk=Monkey
Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 7854:
- ELISA-P
Description
CST's PathScan® Phospho-4E-BP1 (Thr37/Thr46) Sandwich ELISA Antibody Pair is 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 kit are custom formulations specific to kit.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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 37ÂșC.
Background
Translation repressor protein 4E-BP1 (also known as PHAS-1) inhibits cap-dependent translation by binding to the translation initiation factor eIF4E. 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 at 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
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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 Non-phospho-4E-BP1 (Thr46) (87D12) Rabbit mAb
- 9459 Phospho-4E-BP1 (Thr37/46) Antibody
This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.