Affinity purified from rabbit antiserum by affinity chromatography using epitope specific phosphopeptide. The antibody against non phosphopeptide was removed by chromatography using non phosphopeptide corresponding to the phosphorylation site
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/300 - 1/1000. Detects a band of approximately 58 kDa (predicted molecular weight: 58 kDa).
1/50 - 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Receptor-regulated SMAD (R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta (transforming growth factor) and activin type 1 receptor kinases. Binds the TRE element in the promoter region of many genes that are regulated by TGF-beta and, on formation of the SMAD2/SMAD4 complex, activates transcription. May act as a tumor suppressor in colorectal carcinoma.
Expressed at high levels in skeletal muscle, heart and placenta.
Belongs to the dwarfin/SMAD family. Contains 1 MH1 (MAD homology 1) domain. Contains 1 MH2 (MAD homology 2) domain.
Phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases. Able to interact with SMURF2 when phosphorylated on Ser-465/467, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, phosphorylated on Ser-240 by CaMK2. Phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin. In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation. Acetylated on Lys-19 by coactivators in response to TGF-beta signaling, which increases transcriptional activity. Isoform short: Acetylation increases DNA binding activity in vitro and enhances its association with target promoters in vivo. Acetylation in the nucleus by EP300 is enhanced by TGF-beta.
Cytoplasm. Nucleus. Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue labeling Smad2 (phospho S467) with ab53100. The image on the right is blocked with a phosphopeptide.
Western blot - Anti-Smad2 (phospho S467) antibody (ab53100)This image is courtesy of an anonymous Abreview
Anti-Smad2 (phospho S467) antibody (ab53100) at 1/750 dilution (for 16 hours at 4°C) + Mouse brain whole tissue lysate at 18 µg
Secondary An HRP-conjugated Goat anti-rabbit-IgG polyclonal at 1/7000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 58 kDa Observed band size: 58 kDa
ELISA - Anti-Smad2 (phospho S467) antibody (ab53100)This image is courtesy of an anonymous Abreview
ab53100 detecting phospho-Smad2 in Mouse brain samples by indirect ELISA. Samples were blocked with 5% BSA for 1 hour at 20°C and incubated with the primary antibody (1/750) for 4 hours at 20°C. An HRP-conjugated Goat anti-rabbit IgG polyclonal (1/5000) was used as the secondary antibody.
Yang HJ et al. GP73 promotes invasion and metastasis of bladder cancer by regulating the epithelial-mesenchymal transition through the TGF-ß1/Smad2 signalling pathway. J Cell Mol Med22:1650-1665 (2018).
Read more (PubMed: 29349903) »