Phospho-SMAD1 (Ser463/465)/ SMAD5 (Ser463/465)/ SMAD9 (Ser465/467) (D5B10) Rabbit mAb #13820
- WB
- IP
- IF
- F
- ChIP
Supporting Data
REACTIVITY | H M R |
SENSITIVITY | Endogenous |
MW (kDa) | 60 |
Source/Isotype | Rabbit IgG |
Application Key:
- WB-Western Blotting
- IP-Immunoprecipitation
- IF-Immunofluorescence
- F-Flow Cytometry
- ChIP-Chromatin Immunoprecipitation
Species Cross-Reactivity Key:
- H-Human
- M-Mouse
- R-Rat
Product Information
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Immunofluorescence (Immunocytochemistry) | 1:400 - 1:1600 |
Flow Cytometry (Fixed/Permeabilized) | 1:200 - 1:800 |
Chromatin IP | 1:50 |
Storage
For a carrier free (BSA and azide free) version of this product see product #67959.
Protocol
Specificity / Sensitivity
Species Reactivity:
The antigen sequence used to produce this antibody shares 100% sequence homology with the species listed here, but reactivity has not been tested or confirmed to work by CST. Use of this product with these species is not covered under our Product Performance Guarantee.
Species predicted to react based on 100% sequence homology:
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human SMAD1 and SMAD5 protein.
Background
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 superfamily 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 regulate the transcription of target genes (5). MAP kinases and CDKs 8 and 9 are also reported to phosphorylate residues in the linker region of SMAD1, including Ser206. Phosphorylation of SMAD1 at Ser206 recruits Smurf1 to the linker region and leads to the degradation of SMAD1 (6). Phosphorylation at this site also promotes SMAD1 transcriptional activity 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.
Limited Uses
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