Product Name:
Smad3-pT8
Product Number:
ab-pn658
Target Full Name: Mothers against decapentaplegic homologue 3
Target Alias: SMAD3
Product Type Specific: Smad3 phosphosite-specific antibody
Antibody Code: PN658
Antibody Target Type: Phosphosite-specific
Antibody Phosphosite: T8
Protein UniProt: P84022
Protein SigNET: Smad3
Antibody Type: Polyclonal
Antibody Host Species: Rabbit
Antibody Immunogen Source: Synthetic phosphopeptide patterned after human Smad3
Antibody Immunogen Sequence: ILPF(pT)PPIV(βA)C
Antibody Immunogen Description: Corresponds to amino acid residues I4 to V12. T8 phosphorylation inhibits transcriptional and antiproliferative activities. This is one of the major sites of in vivo phosphorylation of Smad3 based on ≥13 mass spectrometry reports recorded in PhosphoSitePlus. Smad3 is known to be phosphorylated at this site in vitro by CDK2, and CDK4.
Production Method: The immunizing peptide was produced by solid phase synthesis on a multipep peptide synthesizer and purified by reverse-phase hplc chromatography. Purity was assessed by analytical hplc and the amino acid sequence confirmed by mass spectrometry analysis. This peptide was coupled to KLH prior to immunization into rabbits. New Zealand White rabbits were subcutaneously injected with KLH-coupled immunizing peptide every 4 weeks for 4 months. The sera from these animals was applied onto an agarose column to which the immunogen peptide was thio-linked. Antibody was eluted from the column with 0.1 M glycine, pH 2.5. Subsequently, the antibody solution was neutralized to pH 7.0 with saturated Tris.
Antibody Modification: Unconjugated. Contact KInexus if you are interest in having the antibody biotinylated or coupled with fluorescent dyes.
Antibody Concentration: 1 mg/ml
Storage Buffer: Phosphate buffered saline pH 7.4, 0.05% Thimerasol
Storage Conditions: For long term storage, keep frozen at -40°C or lower. Stock solution can be kept at +4°C for more than 3 months. Avoid repeated freeze-thaw cycles.
Product Use: Western blotting | Antibody microarray
Antibody Dilution Recommended: 2 µg/ml for immunoblotting
Antibody Species Reactivity: This antibody detects the target phosphoprotein in the following species due to conservation of amino acid sequence: Human | Chimpanzee | Rhesus macaque | Dog | Rat | Mouse | Platypus | Chicken | Frog | Zebra fish | Sea urchin .
Related Product 1: Smad4 Y513 phosphorylation antibody (Cat. No.: AB-PN725)
Scientific Background: Smad3 (Mothers against decapentaplegic homolog 3) a receptor-regulated Smad (R-SMAD) protein that belongs to the dwarfin/SMAD family. It functions as an intracellular signal transducer and transcription factor within the transforming growth factor-beta (TGF-β) signaling pathway. It is activated by TGF-β receptors, and it forms a complex with Smad4 to regulate gene expression, influencing cell proliferation, differentiation, apoptosis, and acting as a tumour suppressor. It transmits signals from the cell surface to the nucleus, binding to specific DNA sequences (TRE elements) to modulate target gene transcription. It functions downstream of TGF-β and activin receptor kinases, regulating cell cycle arrest, extracellular matrix formation, and immune responses. The Smad3/Smad4 complex has an inhibitory effect on wound healing probably by modulating both growth and migration of primary keratinocytes and by altering the TGF-mediated chemotaxis of monocytes. This effect on wound healing appears to be hormone-sensitive. It is a regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. In the mouse, it is required for establishment of the mucosal immune response and proper development of skeleton. It can form complexes that include, for example, Smad4/JUN/FOS, Smad2/Smad4, and AIP1, ACVR2A, and ACVR1B. IT Interacts with DACH1, inhibiting TGF-beta signaling. Interaction (via the MH2 domain) with LEMD3, represses Smad3 transcriptional activity by preventing the formation of the heteromeric complex with Smad4 and translocation to the nucleus. It interacts (via the linker region) with EP300 (C-terminal), which promotes Smad3 acetylation. Interaction with SKI, represses SMAD3 transcriptional activity. Interaction with PPM1A which dephosphorylates Smad3 in the C-terminal SXS motif leads to disruption of the Smad2/3-Smad4 complex, nuclear export and termination of TGF-beta signaling. Interaction with RANBP3 results in the export of dephosphorylated Smad3 out of the nucleus and termination of the TGF-beta signaling. TGF-beta stimulation enhances interactions with DAB2. It interacts with PPP5C, decreasing Smad3 phosphorylation and protein levels. Interaction (when phosphorylated) with RNF111 results in its activation as an enhancer of the transcriptional responses by mediating ubiquitination and degradation of Smad3 inhibitors. Smad3 is involved in embryonic development, wound healing (can act as an inhibitor), and chondrogenesis. Mutations are associated with Loeys-Dietz Syndrome 3 (LDS3) and aneurysms-osteoarthritis syndrome. Dysregulation is linked to fibrosis and cancer. This description may include information annotated by UniProt and/or Google AI.