Revision 3
#7182
Store at +4C
PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit
1 Kit
(96 assays)
Species Cross Reactivity:
H M Mk
UniProt ID:
#Q92934
Entrez-Gene Id:
#572
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Bad Rabbit mAb Coated Microwells | 62838 | 96 tests | +4C | |
| Phospho-Bad (Ser112) Mouse Detection mAb | 14218 | 1 ea | Green (Lyophilized) | +4C |
| Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 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
CST's PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Bad (Ser112) protein. A Bad rabbit mAb has been coated onto the microwells. After incubation with cell lysates, Bad protein (phospho and nonphospho) is captured by the coated antibody. Following extensive washing, a phospho-Bad (Ser112) mouse mAb is added to detect the captured phospho-Bad 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 absorbance for this developed color is proportional to the quantity of phospho-Bad (Ser112) protein.
*Antibodies in kit are custom formulations specific to kit.
Specificity/Sensitivity
CST's PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit #7182 detects endogenous levels of phospho-Bad (Ser112) protein. A significant induction of Bad phosphorylation at Ser112 can be detected in TPA-treated OVCAR8 cells using PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit #7182. However, the level of total Bad protein (phospho and nonphospho) detected by PathScan® Total Bad Sandwich ELISA Kit #7162 remains unchanged (Figure 1). This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Background
Bad is a proapoptotic member of the Bcl-2 family that promotes cell death by displacing Bax from binding to Bcl-2 and Bcl-xL (1,2). Survival factors, such as IL-3, inhibit the apoptotic activity of Bad by activating intracellular signaling pathways that result in the phosphorylation of Bad at Ser112 and Ser136 (2). Phosphorylation at these sites promotes binding of Bad to 14-3-3 proteins to prevent an association between Bad with Bcl-2 and Bcl-xL (2). Akt phosphorylates Bad at Ser136 to promote cell survival (3,4). Bad is phosphorylated at Ser112 both in vivo and in vitro by p90RSK (5,6) and mitochondria-anchored PKA (7). Phosphorylation at Ser155 in the BH3 domain by PKA plays a critical role in blocking the dimerization of Bad and Bcl-xL (8-10).
Background References
- Yang, E. et al. (1995) Cell 80, 285-291.
- Zha, J. et al. (1996) Cell 87, 619-628.
- Datta, S.R. et al. (1997) Cell 91, 231-241.
- Peso, L. et al. (1997) Science 278, 687-689.
- Bonni, A. et al. (1999) Science 286, 1358-1362.
- Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867.
- Harada, H. et al. (1999) Mol. Cell 3, 413-422.
- Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
- Lizcano, J. et al. (2000) Biochem. J. 349, 547-557.
- Datta, S. et al. (2000) Mol. Cell 6, 41-51.
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 G: Goat 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.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
Limited Uses
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