Anti-phospho Smad1/Smad5/Smad8 (Ser463/465)

Mothers against decapentaplegic homolog 1 (UniProt Q15797; also known as BSP-1, hSMAD1, JV4-1, MAD homolog 1, Mothers against DPP homolog 1, Mad-related protein 1, SMAD 1, SMAD family member 1, Smad1, Transforming growth factor-beta-signaling protein 1) is encoded by the SMAD1 (also known as BSP1, MADH1, MADR1) gene (Gene ID 4086) in human. Mammalian SMADs constitute a family of transcription factors that are subdivided into three groups. SMAD1, 2, 3, 5, and 9 (a.k.a. SMAD8) are receptor-regulated SMADs or R-SMADs. SMAD4 is the only known mammalian common mediator SMAD or Co-SMAD and it mediates both the TGF- and BMP signalling pathways. SMAD6 and SMAD7 are inhibitory SMADs or I-SMADs that block the activation of R-SMADs by Co-SMAD via competitive binding. SMADs share a similar structure feature consisting of the conserved N-terminal Mad-homology 1 (MH1; a.a. 12-136 of hSMAD1, a.a. 13-137 of hSMAD2, a.a. 16-140 of hSMAD8) and C-terminal MH2 (a.a. 271-465 of hSMAD1 & hSMAD2, a.a. 273-467 of hSMAD8) domains joined by a non-conserved linker region among SMADs. MH1 domain in R-SMADs and Co-SMADs contains N-terminus nuclear localization signals (NLS), while SMAD4 has nuclear export signals (NES) in this domain. The MH2 domain is responsible for gene transactivation as well as for mediating SMAD-receptor, SMAD-SMAD, SMAD-coactivators and SMAD-corepressors interactions. A TGF- ligand (TGF- for SMAD2/3 and BMP for SMAD1/5/8) initiates signalling by binding and engaging type I and type II receptor serine/threonine kinases on the cell surface, allowing T RII-dependent phosphorylation of T RI kinase domain, which then instigates signalling by phosphorylating the last two serine residues within the conserved C-terminal end Ser-Ser-X-Ser (SSXS) motif of SMAD proteins.

~55 kDa observed. 52.26/52.16/52.71 kDa (human/mouse/rat SMAD1), 52.26/52.17/52.22 kDa (human/mouse/rat SMAD5), and 52.49/48.64/48.99 kDa Uncharacterized bands may be observed in some lysate(s).

Epitope: C-terminus

KLH-conjugated linear peptide corresponding to human SMAD1 C-terminal end sequence phosphorylated on the last two serine residues (S463/S465).

Anti-phospho Smad1/Smad5/Smad8 (Ser463/465) Antibody, Cat. No. AB3848-I, is a highly specific rabbit polyclonal antibody that targets Smad1//5/8 phosphorylation and has been tested in Immunohistochemistry and Western Blotting.

Immunohistochemistry Analysis: A 1:50 dilution from a representative lot detected Smad1/5/8 phosphorylation in mouse & rat embryo tissue sections.

Research Category
Epigenetics & Nuclear Function

This rabbit polyclonal antibody recognizes SMAD1/5/8 phosphorylated on their last two serine residues within the C-terminal end Ser-Ser-X-Ser (SSXS) motif. Target serine residues are present in human/mouse SMAD1 (S463/S465), rat SMAD1 (S466/S468), human/mouse/rat SMAD5 (S463/S465), human SMAD9 (S465/S467), mouse SMAD9 (S428/S430), and rat SMAD9 (S432/S434), but absent in human SMAD1 spliced isoform 2. SMAD9 is also known as SMAD8. Target bands detection was blocked by the immunogen phosphopeptide, but not by the corresponding non-phosphopeptide.

Affinity purified.

Purified rabbit polyclonal antibdoy in buffer containing 0.1 M Tris-Glycine (pH 7.4) 150 mM NaCl with 0.05% sodium azide

Stable for 1 year at 2-8°C from date of receipt.

Evaluated by Western Blotting in HEK293 cell lysates.

Western Blotting Analysis: A 1:500 dilution of this antibody detected an increased SMAD1/5/8 phosphorylation in 10 µg lysate from BMP2-treated HEK293 cells. Target bands detection was blocked by the immunogen phosphopeptide, but not the corresponding non-phosphopeptide.

Concentration: Please refer to lot specific datasheet.

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