Application

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 CategoryEpigenetics & Nuclear Function

Disclaimer

Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

General description

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.

Immunogen

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

Epitope: C-terminus

Other Notes

Concentration: Please refer to lot specific datasheet.

Physical form

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

Affinity purified.

Quality

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.

Specificity

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.

Storage and Stability

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

Target description

~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).