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9105
Phospho-Bad Antibody Sampler Kit

Phospho-Bad Antibody Sampler Kit #9105

Western Blotting Image 1

Western blot analysis of extracts from COS cells, untreated or TPA-treated, using Phospho-Bad (Ser112) (40A9) Rabbit mAb (upper) or Bad Antibody #9292 (lower).

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Western Blotting Image 2

Western blot analysis of extracts from ACHN cells, untreated or treated with Human Epidermal Growth Factor #8916 (100 ng/ml, 30 min), and C2C12 cells, untreated or insulin-treated (100 nM, 30 min), using Phospho-Bad (Ser136) (D25H8) Rabbit mAb (upper) or Bad (D24A9) Rabbit mAb #9239 (lower).

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Western Blotting Image 3

Western blot analysis of cell extracts from 293 cells transfected with Wild-type Bad, Bad (S112A/S136A), Bad (S155A) and treated with forskolin (30 µM), using Phospho-Bad (Ser155) Antibody (upper) and Bad Antibody #9292 (lower).

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Western Blotting Image 4

Western blot analysis of extracts from various cell lines using Bad (D24A9) Rabbit mAb.

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Western Blotting Image 5

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.

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IHC-P (paraffin) Image 6

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, untreated (left) or lambda phosphatase treated (right), using Phospho-Bad (Ser 112) (40A9) Rabbit mAb.

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Western Blotting Image 7

Western blot analysis of extracts from COS-7 cells, treated with Human Epidermal Growth Factor #8916 (100 ng/ml, 30 min) in the presence or absence of calf intestinal phosphatase (CIP) plus lambda-phosphatase, using Phospho-Bad (Ser136) (D25H8) Rabbit mAb (upper) or Bad (D24A9) Rabbit mAb #9239 (lower).

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IHC-P (paraffin) Image 8

Immunohistochemical analysis of paraffin embedded COS cells untreated (left) or TPA-treated (right), showing induced cytoplasmic staining using Phospho-Bad (Ser112) (40A9) Rabbit mAb.

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Western Blotting Image 9

Western blot analysis of lysates containing Bad Control Proteins #9293, comprised of nonphosphorylated and phosphorylated Bad peptides, using Phospho-Bad (Ser136) (D25H8) Rabbit mAb.

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IHC-P (paraffin) Image 10

Immunohistochemical analysis of paraffin-embedded human prostate carcinoma, using Phospho-Bad (Ser 112) (40A9) Rabbit Monoclonal Antibody preincubated with control peptide (left) or Phospho-Bad (Ser 112) Blocking Peptide (IHC Specific) #1026 (right).

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IHC-P (paraffin) Image 11

Immunohistochemical analysis of paraffin-embedded Non-Hodgkin's lymphoma, using Phospho-Bad (Ser112) (40A9) Rabbit mAb.

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Flow Cytometry Image 12

Flow cytometric analysis of COS cells, untreated (blue) or TPA/Calyculin A treated (green), using Phospho-Bad (Ser112) (40A9) Rabbit mAb.

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Product Includes Quantity Applications Reactivity MW(kDa) Isotype
Phospho-Bad (Ser112) (40A9) Rabbit mAb 5284 20 µl
  • WB
  • IHC
  • F
H M R Mk 23 Rabbit IgG
Phospho-Bad (Ser136) (D25H8) Rabbit mAb 4366 20 µl
  • WB
  • IP
H M Mk 23 Rabbit IgG
Phospho-Bad (Ser155) Antibody 9297 20 µl
  • WB
M 23 Rabbit 
Bad (D24A9) Rabbit mAb 9239 20 µl
  • WB
H M R Mk 23 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Phospho-Bad Antibody Sampler Kit provides an economical means to investigate the role of Bad protein in apoptosis. The kit contains primary and secondary antibodies to perform two Western blots with each antibody.

Bad (D24A9) Rabbit mAb detects endogenous levels of total Bad protein. Phospho-Bad (Ser112) (40A9) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser112 (equivalent to Ser75 in human and Ser113 in rat). Phospho-Bad (Ser136) (D25H8) Rabbit mAb detects endogenous levels of Bad only when phosphorylated at Ser136 (equivalent to Ser99 in human and Ser137 in rat). Phospho-Bad (Ser155) Antibody detects transfected levels of Bad only when phosphorylated at Ser155 (equivalent to Ser118 in human and Ser156 in rat).

Bad (D24A9) Rabbit mAb (#9239) was prepared by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro102 of human Bad protein. Phospho-specific antibodies (#4366, #5284, #9297) were prepared by respectively immunizing animals with synthetic phosphopeptides corresponding to residues surrounding Ser112, Ser136, Ser155 of mouse Bad protein. Polyclonal Phospho-Bad (Ser155) Antibody (#9297) was purified by protein A and peptide affinity chromatography.

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, 619-28.
  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.
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.

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