ARG66276

anti-Smad 3 phospho (Thr179) antibody

anti-Smad 3 phospho (Thr179) antibody for Western blot and Human,Mouse,Rat

Overview

Product Description Rabbit Polyclonal antibody recognizes Smad 3 phospho (Thr179)
Tested Reactivity Hu, Ms, Rat
Tested Application WB
Specificity The antibody detects endogenous level of Smad3 only when phosphorylated at threonine 179.
Host Rabbit
Clonality Polyclonal
Isotype IgG
Target Name Smad 3
Antigen Species Human
Immunogen KLH-conjugated phospho-specific peptide around Thr179 (PET(p)PP) of Human Smad3.
Conjugation Un-conjugated
Alternate Names JV15-2; SMAD 3; MADH3; Mothers against decapentaplegic homolog 3; LDS3; Smad3; HsT17436; Mothers against DPP homolog 3; hSMAD3; Mad3; HSPC193; hMAD-3; SMAD family member 3; LDS1C; MAD homolog 3

Application Instructions

Application Suggestion
Tested Application Dilution
WB1:500 - 1:1000
Application Note * The dilutions indicate recommended starting dilutions and the optimal dilutions or concentrations should be determined by the scientist.

Properties

Form Liquid
Purification Affinity purification with phospho-specific peptide and the non-phospho specific antibodies were removed by chromatography using non-phosphopeptide.
Buffer PBS (pH 7.4), 150mM NaCl, 0.02% Sodium azide and 50% Glycerol.
Preservative 0.02% Sodium azide
Stabilizer 50% Glycerol
Concentration 1 mg/ml
Storage Instruction For continuous use, store undiluted antibody at 2-8°C for up to a week. For long-term storage, aliquot and store at -20°C. Storage in frost free freezers is not recommended. Avoid repeated freeze/thaw cycles. Suggest spin the vial prior to opening. The antibody solution should be gently mixed before use.
Note For laboratory research only, not for drug, diagnostic or other use.

Bioinformation

Database Links

GeneID: 17127 Mouse SMAD3

GeneID: 25631 Rat SMAD3

GeneID: 4088 Human SMAD3

Gene Symbol SMAD3
Gene Full Name SMAD family member 3
Background The protein encoded by this gene belongs to the SMAD, a family of proteins similar to the gene products of the Drosophila gene 'mothers against decapentaplegic' (Mad) and the C. elegans gene Sma. SMAD proteins are signal transducers and transcriptional modulators that mediate multiple signaling pathways. This protein functions as a transcriptional modulator activated by transforming growth factor-beta and is thought to play a role in the regulation of carcinogenesis. [provided by RefSeq, Apr 2009]
Function 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 SMAD3/SMAD4 complex, activates transcription. Also can form a SMAD3/SMAD4/JUN/FOS complex at the AP-1/SMAD site to regulate TGF-beta-mediated transcription. 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. Regulator of chondrogenesis and osteogenesis and inhibits early healing of bone fractures. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator. [UniProt]
Calculated MW 48 kDa
PTM Phosphorylated on serine and threonine residues. Enhanced phosphorylation in the linker region on Thr-179, Ser-204 and Ser-208 on EGF and TGF-beta treatment. Ser-208 is the main site of MAPK-mediated phosphorylation. CDK-mediated phosphorylation occurs in a cell-cycle dependent manner and inhibits both the transcriptional activity and antiproliferative functions of SMAD3. This phosphorylation is inhibited by flavopiridol. Maximum phosphorylation at the G(1)/S junction. Also phosphorylated on serine residues in the C-terminal SXS motif by TGFBR1 and ACVR1. TGFBR1-mediated phosphorylation at these C-terminal sites is required for interaction with SMAD4, nuclear location and transactivational activity, and appears to be a prerequisite for the TGF-beta mediated phosphorylation in the linker region. Dephosphorylated in the C-terminal SXS motif by PPM1A. This dephosphorylation disrupts the interaction with SMAD4, promotes nuclear export and terminates TGF-beta-mediated signaling. Phosphorylation at Ser-418 by CSNK1G2/CK1 promotes ligand-dependent ubiquitination and subsequent proteasome degradation, thus inhibiting SMAD3-mediated TGF-beta responses. Phosphorylated by PDPK1.

Acetylation in the nucleus by EP300 in the MH2 domain regulates positively its transcriptional activity and is enhanced by TGF-beta.

Poly-ADP-ribosylated by PARP1 and PARP2. ADP-ribosylation negatively regulates SMAD3 transcriptional responses during the course of TGF-beta signaling.

Ubiquitinated. Monoubiquitinated, leading to prevent DNA-binding (PubMed:21947082). Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes (PubMed:21947082). Ubiquitinated by RNF111, leading to its degradation: only SMAD3 proteins that are 'in use' are targeted by RNF111, RNF111 playing a key role in activating SMAD3 and regulating its turnover (By similarity). Undergoes STUB1-mediated ubiquitination and degradation (PubMed:24613385). [UniProt]

Images (1) Click the Picture to Zoom In

  • ARG66276 anti-Smad 3 phospho (Thr179) antibody WB image

    Western blot: HeLa cells treated with TNF and stained with ARG66276 anti-Smad 3 phospho (Thr179) antibody. The lane on the right is treated with the antigen-specific peptide.