Product Pathways - PathScan ELISA
PathScan® Total Bad Sandwich ELISA Antibody Pair #7840
|7840S||1 Kit (Reagents for 4 x 96 well plates)||---||In Stock||---|
|7840||carrier free and custom formulation / quantity||email request|
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|Kit Includes||Volume||Cap Color|
|Bad Capture Ab (100X)||0.4 ml||Pink|
|Bad Detection Ab (100X)||0.4 ml||Blue|
|Anti-mouse IgG, HRP-linked Antibody||0.04 ml||Yellow|
Storage: Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Important Ordering Details
Product is assembled upon order to ensure maximum activity. United States: Please allow up to two weeks for your order to be processed and shipped. Outside of the United States: Please allow up to three weeks, depending on the country, for your order to be processed and shipped.
CST's PathScan® Total Bad Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total Bad Sandwich ELISA Kit #7162. 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 Bad Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Bad Detection Antibody and an 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 total Bad protein.
*Antibodies in this kit are custom formulations specific to the 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.
The relationship between the protein concentration of lysates from untreated and TPA-treated OVCAR8 cells and the absorbance at 450 nm using PathScan® Total Bad Sandwich ELISA Antibody Pair #7840 is shown. Cells were serum starved overnight and then treated with 200 nm TPA for 30 min. at 37ºC.
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).
- 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.
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- 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
- 7162 PathScan® Total Bad Sandwich ELISA Kit
- 7182 PathScan® Phospho-Bad (Ser112) Sandwich ELISA Kit
- 7842 PathScan® Phospho-Bad (Ser112) Sandwich ELISA Antibody Pair
- 9292 Bad Antibody
- 9268 Bad (11E3) Rabbit mAb (IP Preferred)
For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.