Product Pathways - TGF-beta/Smad Signaling
Smad1 (D59D7) XP® Rabbit mAb #6944
|6944S||100 µl (10 western blots)||---||In Stock||---|
|6944P||40 µl (4 western blots)||---||In Stock||---|
|6944||carrier free and custom formulation / quantity||email request|
|W||1:1000||Human, Mouse, Monkey||Endogenous||60||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry, ChIP=Chromatin IP
Species predicted to react based on 100% sequence homology: Xenopus, Bovine.
Specificity / Sensitivity
Smad1 (D59D7) XP® Rabbit mAb recognizes endogenous levels of total Smad1 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser190 of human Smad1 protein.
Western blot analysis of extracts from various cell lines using Smad1 (D59D7) XP® Rabbit mAb.
Confocal immunofluorescent analysis of HT-1080 cells, untreated (left) or treated with human BMP2 #4697 (right), using Smad1 (D59D7) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).
Flow cytometric analysis of HT-1080 cells using Smad1 (D59D7) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).
Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 MCF7 cells treated with Human BMP2 #4697 (50 ng/ml) for one hour and either 5 μl of Smad1 (D59D7) XP® 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 Smad9 (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. (1996) Genes Dev 10, 1580-94.
- Hoodless, P.A. et al. (1996) Cell 85, 489-500.
- Klemm, J.D. et al. (1998) Annu Rev Immunol 16, 569-92.
- Kretzschmar, M. et al. (1997) Genes Dev 11, 984-95.
- Whitman, M. (1998) Genes Dev 12, 2445-62.
- Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
- Alarcón, C. et al. (2009) Cell 139, 757-69.
- Katsu, K. et al. (2013) Dev Biol 374, 71-84. Applications: Western Blotting.
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
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This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
XP is a registered trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.