Product Pathways - TGF-beta/Smad Signaling
Smad5 (D4G2) Rabbit mAb #12534
|12534S||100 µl (10 western blots)||---||In Stock||---|
|12534||carrier free and custom formulation / quantity||email request|
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|W||1:1000||Human, Mouse, Rat, Monkey||Endogenous||60||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP
Specificity / Sensitivity
Smad5 (D4G2) Rabbit mAb recognizes endogenous levels of total Smad5 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro249 of human Smad5 protein.
Western blot analysis of extracts from various cell lines using Smad5 (D4G2) Rabbit mAb.
Immunoprecipitation of Smad5 from HT-1080 cell extracts using Normal Rabbit IgG #2729 (lane 2) or Smad5 (D4G2) Rabbit mAb (lane 3). Lane 1 is 10% input. Western blot analysis was performed using Smad5 (D4G2) Rabbit mAb.
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with Human BMP2 #4697 (50 ng/ml, 1 hr) and either 5 μl of Smad5 (D4G2) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human ID1 Promoter Primers #5139, human Smad6 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
Bone morphogenetic proteins (BMPs) constitute a large family of signaling molecules that regulate a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation, and apoptosis (1,2). BMP receptors are members of the TGF-β family of Ser/Thr kinase receptors. Ligand binding induces multimerization, autophosphorylation, and activation of these receptors (3-5). They subsequently phosphorylate Smad1 at Ser463 and Ser465 in the carboxy-terminal motif SSXS, as well as Smad5 and Smad8 at their corresponding sites. These phosphorylated Smads dimerize with the coactivating Smad4 and translocate to the nucleus, where they stimulate transcription of target genes (5).
MAP kinases and CDKs 8 and 9 phosphorylate residues in the linker region of Smad1, including Ser206. The phosphorylation of Ser206 recruits Smurf1 to the linker region and leads to the degradation of Smad1 (6). Phosphorylation of this site also promotes Smad1 transcriptional action by recruiting YAP to the linker region (7).
- Hogan, B.L. et al. (1996) Genes Dev. 10, 1580-1594.
- Hoodless, P.A. et al. (1996) Cell 85, 489-500.
- Klemm, J.D. et al. (1998) Annu. Rev. Immunol. 16, 569-592.
- Kretzschmar, M. et al. (1997) Genes Dev. 11, 984-995.
- Whitman, M. (1998) Genes Dev. 12, 2445-2462.
- Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
- Alarcón, C. et al. (2009) Cell 139, 757-69.
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For Research Use Only. Not For Use In Diagnostic Procedures.
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