Smad1 Antibody

9743:

ChIP Agarose , ChIP Magnetic , Immunoprecipitation , Western Blotting

Smad1 Antibody detects endogenous levels of total Smad1 protein. No cross reactivity was observed with other family members.

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser190 of human Smad1. Antibodies were purified by protein A and peptide affinity chromatography.

Western Blotting

Western blot analysis of extracts from various cell lines using Smad1 Antibody.

Chromatin IP

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 20 μl of Smad1 Antibody 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).

1. Hogan, B.L. et al. (1996) Genes Dev. 10, 1580-1594.
2. Hoodless, P.A. et al. (1996) Cell 85, 489-500.
3. Klemm, J.D. et al. (1998) Annu. Rev. Immunol. 16, 569-592.
4. Kretzschmar, M. et al. (1997) Genes Dev. 11, 984-995.
5. Whitman, M. (1998) Genes Dev. 12, 2445-2462.
6. Sapkota, G. et al. (2007) Mol Cell 25, 441-54.
7. Alarcón, C. et al. (2009) Cell 139, 757-69.

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• 6944 Smad1 (D59D7) XP^® Rabbit mAb
• 5753 Phospho-Smad1 (Ser206) (D40B7) Rabbit mAb
• 9511 Phospho-Smad1 (Ser463/465)/ Smad5 (Ser463/465)/ Smad8 (Ser426/428) Antibody
• 9516 Phospho-Smad1/5 (Ser463/465) (41D10) Rabbit mAb
• 9963 Phospho-Smad Antibody Sampler Kit
• 7071 Phototope^® -HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7720 Prestained Protein Marker, Broad Range (Premixed Format)
• 7727 Biotinylated Protein Ladder Detection Pack
• 7003 20X LumiGLO^® Reagent and 20X Peroxide

For Research Use Only. Not For Use In Diagnostic Procedures.