Rabbit Smad2 Polyclonal Antibody | anti-Smad2 antibody

Phospho-Smad2 (Ser465+Ser467) Antibody

Cat. Num. AAA9614315

• Gene Names: SMAD2; JV18; MADH2; MADR2; JV18-1; hMAD-2; hSMAD2
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Zebrafish (100%), Bovine (100%), Sheep (100%), Rabbit (100%), Dog (100%), Xenopus (100%)
• Applications: Western Blot, Immunohistochemistry, Immunofluorescence, Immunocytochemistry, ELISA
• Purity: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Synonyms: Smad2; Polyclonal Antibody; Phospho-Smad2 (Ser465+Ser467) Antibody; Drosophila; homolog of; MADR2; hMAD-2; HsMAD2; JV18; JV18-1; JV181; MAD; MAD homolog 2; MAD Related Protein 2; Mad-related protein 2; MADH2; MGC22139; MGC34440; Mother against DPP homolog 2; Mothers against decapentaplegic homolog 2; Mothers against decapentaplegic; 2; Mothers against DPP homolog 2; OTTHUMP00000163489; Sma and Mad related protein 2; Sma- and Mad-related protein 2 MAD; SMAD 2; SMAD family member 2; SMAD; mothers against DPP homolog 2; SMAD2; SMAD2_HUMAN; anti-Smad2 antibody

• Host: Rabbit
• Reactivity: Human, Mouse, Rat; Predicted Reactivity: Pig (100%), Zebrafish (100%), Bovine (100%), Sheep (100%), Rabbit (100%), Dog (100%), Xenopus (100%)
• Clonality: Polyclonal
• Isotype: Rabbit IgG
• Specificity: Phospho-Smad2 (Ser465+Ser467) Antibody detects endogenous levels of Smad2 only when phosphorylated at Ser465+Ser467.; Tissue Specificity: Expressed at high levels in skeletal muscle, endothelial cells, heart and placenta.
• Purity/Purification: The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.
• Form/Format: Liquid. Rabbit IgG in phosphate buffered saline, pH7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
• Concentration: 1mg/ml (varies by lot)

• Applicable Applications for anti-Smad2 antibody: Western Blot (WB), Immunohistochemistry (IHC), Immunofluorescence (IF), Immunocytochemistry (ICC), Peptide ELISA (EIA)
• Application Notes: WB: 1:1000-3000; IF/ICC: 1:100-1:500; IHC: 1:50-1:200; Peptide ELISA: 1:20,000-1:40,000
• Immunogen: A synthesized peptide derived from human Smad2 around the phosphorylation site of Ser465+Ser467.
• Conjugation: Unconjugated
• Fragment: Fab fragment
• Post Translational Modifications: 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. TGF-beta-induced Ser-465/467 phosphorylation declines progressively in a KMT5A-dependent manner. 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. Phosphorylated by PDPK1.In response to TGF-beta, ubiquitinated by NEDD4L; which promotes its degradation. Monoubiquitinated, leading to prevent DNA-binding (By similarity). Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes. Ubiquitinated by RNF111, leading to its degradation: only SMAD2 proteins that are 'in use' are targeted by RNF111, RNF111 playing a key role in activating SMAD2 and regulating its turnover (By similarity). 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.
• Subunit Structure: Monomer; the absence of TGF-beta. Heterodimer; in the presence of TGF-beta. Forms a heterodimer with co-SMAD, SMAD4, in the nucleus to form the transactivation complex SMAD2/SMAD4. Interacts with AIP1, HGS, PML and WWP1 (By similarity). Interacts with NEDD4L in response to TGF-beta (By similarity). Found in a complex with SMAD3 and TRIM33 upon addition of TGF-beta. Interacts with ACVR1B, SMAD3 and TRIM33. Interacts (via the MH2 domain) with ZFYVE9; may form trimers with the SMAD4 co-SMAD. Interacts with FOXH1, homeobox protein TGIF, PEBP2-alpha subunit, CREB-binding protein (CBP), EP300, SKI and SNW1. Interacts with SNON; when phosphorylated at Ser-465/467. Interacts with SKOR1 and SKOR2. Interacts with PRDM16. Interacts (via MH2 domain) with LEMD3. Interacts with RBPMS. Interacts with WWP1. Interacts (dephosphorylated form, via the MH1 and MH2 domains) with RANBP3 (via its C-terminal R domain); the interaction results in the export of dephosphorylated SMAD3 out of the nucleus and termination of the TGF-beta signaling. Interacts with PDPK1 (via pHdomain). Interacts with DAB2; the interactions are enhanced upon TGF-beta stimulation. Interacts with USP15. Interacts with PPP5C. Interacts with ZNF580. Interacts with LDLRAD4 (via the SMAD interaction motif). Interacts (via MH2 domain) with PMEPA1 (via the SMAD interaction motif). Interacts with ZFHX3. Interacts with ZNF451. Identified in a complex that contains at least ZNF451, SMAD2, SMAD3 and SMAD4. Interacts weakly with ZNF8 (By similarity). Interacts (when phosphorylated) with RNF111; RNF111 acts as an enhancer of the transcriptional responses by mediating ubiquitination and degradation of SMAD2 inhibitors (By similarity).
• Similarity: Belongs to the dwarfin/SMAD family.
• Subcellular Location: Cytoplasm. Nucleus.; Note: Cytoplasmic and nuclear in the absence of TGF-beta. On TGF-beta stimulation, migrates to the nucleus when complexed with SMAD4 (PubMed:9865696). On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1 (PubMed:16751101, PubMed:19289081).
• Preparation and Storage: Store at -20 degree C. Stable for 12 months from date of receipt.

Western Blot (WB)

(Western blot analysis Smad2 (Phospho-Ser465+Ser467) using PMA treated NIH-3T3 whole cell lysates.-/+ means absence or presence of N peptide（non-phospho peptide) and P peptide(phospho peptide).)

Immunohistochemistry (IHC)

(Staining Rat stomach tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human lung cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human mammary cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Human normal tissues adjacent to colorectal cancer by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Rat spleen tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Mouse brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Mouse testis tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Immunohistochemistry (IHC)

(At 1/100 staining Rat brain tissue by IHC-P. The sample was formaldehyde fixed and a heat mediated antigen retrieval step in citrate buffer was performed. The sample was then blocked and incubated with the primary antibody at 4 degree C overnight. An HRP conjugated anti-Rabbit antibody was used as the secondary antibody.)

Testing Data

(At 25 degree C. Samples were then incubated with primary Ab(At 37 degree C. An AlexaFluor594 conjugated goat anti-rabbit IgG(H+L) Ab(Red) and an AlexaFluor488 conjugated goat anti-mouse IgG(H+L) Ab(Green) were used as the secondary antibody.The nuclear counter stain is DAPI(blue).)

Related Product Information for anti-Smad2 antibody

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. Positively regulates PDPK1 kinase activity by stimulating its dissociation from the 14-3-3 protein YWHAQ which acts as a negative regulator.

NCBI and Uniprot Product Information

• NCBI GI #: 51173730
• NCBI GeneID: 4087
• NCBI Accession #: NP_001003652.1
• NCBI GenBank Nucleotide #: NM_001003652.3
• UniProt Accession #: Q15796
• Molecular Weight: 52,306 Da
• NCBI Official Full Name: mothers against decapentaplegic homolog 2 isoform 1
• NCBI Official Synonym Full Names: SMAD family member 2
• NCBI Official Symbol: SMAD2
• NCBI Official Synonym Symbols: JV18; MADH2; MADR2; JV18-1; hMAD-2; hSMAD2
• NCBI Protein Information: mothers against decapentaplegic homolog 2; MAD homolog 2; mother against DPP homolog 2; Sma- and Mad-related protein 2; SMAD, mothers against DPP homolog 2
• UniProt Protein Name: Mothers against decapentaplegic homolog 2
• Protein Family: Mothers against decapentaplegic
• UniProt Gene Name: SMAD2
• UniProt Synonym Gene Names: MADH2; MADR2; MAD homolog 2; Mothers against DPP homolog 2; hMAD-2; SMAD 2; Smad2; hSMAD2
• UniProt Entry Name: SMAD2_HUMAN

NCBI Description

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 mediates the signal of the transforming growth factor (TGF)-beta, and thus regulates multiple cellular processes, such as cell proliferation, apoptosis, and differentiation. This protein is recruited to the TGF-beta receptors through its interaction with the SMAD anchor for receptor activation (SARA) protein. In response to TGF-beta signal, this protein is phosphorylated by the TGF-beta receptors. The phosphorylation induces the dissociation of this protein with SARA and the association with the family member SMAD4. The association with SMAD4 is important for the translocation of this protein into the nucleus, where it binds to target promoters and forms a transcription repressor complex with other cofactors. This protein can also be phosphorylated by activin type 1 receptor kinase, and mediates the signal from the activin. Alternatively spliced transcript variants have been observed for this gene. [provided by RefSeq, May 2012]

Uniprot Description

SMAD2: ubiquitously expressed transcription factor phosphorylated and activated by TGF-beta receptor-type kinases. Participates in a wide range of critical processes including morphogenesis, cell-fate determination, proliferation, differentiation and apoptosis. Phosphorylated Smads dimerize with collaborating Smad4 and are translocated into the nucleus, where the transcription of target genes is stimulated. Two alternatively spliced isoforms have been described.

Protein type: DNA-binding; Transcription factor

Chromosomal Location of Human Ortholog: 18q21.1

Cellular Component: nucleoplasm; transcription factor complex; cytoplasm; nuclear chromatin; nucleus; cytosol

Molecular Function: transcription activator binding; metal ion binding; transcription factor binding; transforming growth factor beta receptor, pathway-specific cytoplasmic mediator activity; phosphatase binding; protein binding; DNA binding; ubiquitin protein ligase binding; double-stranded DNA binding; chromatin binding; SMAD binding; transcription factor activity; transforming growth factor beta receptor binding

Biological Process: developmental growth; somatic stem cell maintenance; positive regulation of transcription, DNA-dependent; activin receptor signaling pathway; paraxial mesoderm morphogenesis; gastrulation; primary microRNA processing; negative regulation of transcription from RNA polymerase II promoter; palate development; post-embryonic development; anterior/posterior pattern formation; negative regulation of cell proliferation; regulation of transforming growth factor beta receptor signaling pathway; ureteric bud development; transforming growth factor beta receptor signaling pathway; pancreas development; embryonic foregut morphogenesis; response to glucose stimulus; positive regulation of BMP signaling pathway; pericardium development; transcription initiation from RNA polymerase II promoter; cell fate commitment; regulation of binding; transcription, DNA-dependent; in utero embryonic development; common-partner SMAD protein phosphorylation; embryonic cranial skeleton morphogenesis; SMAD protein complex assembly; organ growth; zygotic determination of dorsal/ventral axis; mesoderm formation; endoderm formation; insulin secretion; gene expression; positive regulation of transcription from RNA polymerase II promoter; negative regulation of transcription, DNA-dependent; negative regulation of transforming growth factor beta receptor signaling pathway; lung development