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9290
PhosphoPlus® Bad (Ser112/136) Antibody Kit
Primary Antibodies
PhosphoPlus Antibody Kit

PhosphoPlus® Bad (Ser112/136) Antibody Kit #9290

Citations (0)
PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 1

pEBG-mBad

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 2

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Bad siRNA I (+), using Bad Antibody and p42 MAPK Antibody #9108. The Bad antibody confirms silencing of Bad expression, while the p42 MAPK antibody is used to control for loading and specificity of Bad siRNA.

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 3

Flow cytometric analysis of COS cells, untreated (blue) or Calyculin A/TPA-treated (green), usingPhospho-Bad (Ser112) Antibody compared to Rabbit (DA1E) mAb IgG Isotype Control #3900 (red).

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 4

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 5

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Fluorescein Conjugate) #6201 (-) or SignalSilence® Bad siRNA II (+), using Bad Antibody and α-Tubulin (11H10) Rabbit mAb #2125. The Bad antibody confirms silencing of Bad expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of Bad siRNA.

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 6

Western blot analysis of extracts from 293 cells transfected with Wild-type Bad, Bad (Ser112A), Bad (S136A), Bad (S112A/S136A) and treated with TPA or forskolin, using Phospho-Bad (Ser112) Antibody (upper) or Bad Antibody #9292 (lower).

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 7

Western blot analysis of GST-Bad, phosphorylated by CKII or PKA in vitro, using Phospho-Bad (Ser112) Antibody #9291 (top), Phospho-Bad (Ser136) Antibody (middle) or Bad Antibody #9292 (bottom).

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 8

Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Bad Antibody (right) or Phospho-Bad (Ser112) Antibody #9291 (left).

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 9

Western blot analysis of extracts from 293 cells transfected with Wild-type Bad, Bad (Ser112A), Bad (S136A) or Bad (S112A/S136A), untreated, TPA-treated or forskolin-treated, using Phospho-Bad (Ser112) Antibody #9291 (top), Phospho-Bad (Ser136) Antibody #9295 (middle) or Bad Antibody (bottom).

PhosphoPlus® Bad (Ser112/136) Antibody Kit: Image 10

Western blot analysis of Bad fusion protein phosphorylated by CKII or PKA in vitro, using Phospho-Bad (Ser112) Antibody (upper) or Bad Antibody #9292 (lower).

To Purchase # 9290

Specificity / Sensitivity

Phospho-Bad (Ser112) Antibody and Phospho-Bad (Ser136) Antibody detect Bad only when phosphorylated at serine 112 or serine 136, respectively. Bad antibody detects endogenous levels of total Bad protein. These antibodies do not cross-react with related family members. It is necessary to overexpress Bad to detect Ser136 phosphorylation.

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).

  1. Yang, E. et al. (1995) Cell 80, 285-291.
  2. Zha, J. et al. (1996) Cell 87, 619-628.
  3. Datta, S.R. et al. (1997) Cell 91, 231-241.
  4. Peso, L. et al. (1997) Science 278, 687-689.
  5. Bonni, A. et al. (1999) Science 286, 1358-1362.
  6. Tan, Y. et al. (1999) J. Biol. Chem. 274, 34859-34867.
  7. Harada, H. et al. (1999) Mol. Cell 3, 413-422.
  8. Tan, Y. et al. (2000) J. Biol. Chem. 275, 25865-25869.
  9. Lizcano, J. et al. (2000) Biochem. J. 349, 547-557.
  10. Datta, S. et al. (2000) Mol. Cell 6, 41-51.

Pathways & Proteins

Explore pathways + proteins related to this product.

Limited Uses

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Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus is a trademark of Cell Signaling Technology, Inc.
LumiGLO is a registered trademark of Kirkegaard & Perry Laboratories.