WB, W-S, IF-IC, FC-FP
H M R
Endogenous
60
Rabbit
#Q99717, #Q15797
4090, 4086
Product Information
Product Usage Information
| Application | Dilution |
|---|---|
| Western Blotting | 1:1000 |
| Simple Western™ | 1:10 - 1:50 |
| Immunofluorescence (Immunocytochemistry) | 1:800 |
| Flow Cytometry (Fixed/Permeabilized) | 1:400 - 1:1600 |
Storage
For a carrier free (BSA and azide free) version of this product see product #52937.
Specificity / Sensitivity
Species Reactivity:
Human, Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser463/465 of human SMAD5.
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.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
WB: Western Blotting W-S: Simple Western™ IF-IC: Immunofluorescence (Immunocytochemistry) FC-FP: Flow Cytometry (Fixed/Permeabilized)
Cross-Reactivity Key
H: human M: mouse R: rat Hm: hamster Mk: monkey Vir: virus Mi: mink C: chicken Dm: D. melanogaster X: Xenopus Z: zebrafish B: bovine Dg: dog Pg: pig Sc: S. cerevisiae Ce: C. elegans Hr: horse GP: Guinea Pig Rab: rabbit All: all species expected
Trademarks and Patents
Limited Uses
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