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  • Product Name
    Phospho-Smad2 (Ser465+Ser467) Antibody
  • Catalog No.
    AF8314
  • RRID
    AB_2840376
  • Source
    Rabbit
  • Application
    WB,IHC,IF/ICC,ELISA
  • Reactivity
    Human, Mouse, Rat
  • Prediction
    Pig(100%), Zebrafish(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(100%)
  • UniProt
  • Mol.Wt
    (Observed)58kD; (Calculated)52kDa
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

Drosophila, homolog of, MADR2; hMAD-2; HsMAD2; JV18; JV18-1; JV181; MAD; MAD homolog 2; MAD Related Protein 2; Mad-related protein 2; MADH2; MADR2; MGC22139; MGC34440; Mother against DPP homolog 2; Mothers against decapentaplegic homolog 2; Mothers against decapentaplegic, Drosophila, homolog of, 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;

Applications:

WB 1:1000-3000, IF/ICC 1:100-1:500, IHC 1:50-1:200, ELISA(peptide) 1:20000-1:40000

Reactivity:

Human, Mouse, Rat

Predicted Reactivity:

Pig(100%), Zebrafish(100%), Bovine(100%), Sheep(100%), Rabbit(100%), Dog(100%), Xenopus(100%)

Source:

Rabbit

Clonality:

Polyclonal

Purification:

The antibody is from purified rabbit serum by affinity purification via sequential chromatography on phospho-peptide and non-phospho-peptide affinity columns.

Specificity:

Phospho-Smad2 (Ser465+Ser467) Antibody detects endogenous levels of Smad2 only when phosphorylated at Ser465+Ser467.

RRID:

AB_2840376
Please cite this product as: Affinity Biosciences Cat# AF8314, RRID:AB_2840376.

Format:

Liquid

Concentration:

1mg/ml

Storage Condition and Buffer:

Rabbit IgG in phosphate buffered saline , pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.Store at -20 °C.Stable for 12 months from date of receipt.

Immunogen Information

Immunogen:

A synthesized peptide derived from human Smad2 around the phosphorylation site of Ser465+Ser467.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

SMAD2

Molecular Weight:

Observed Mol.Wt.: (Observed)58kD.
Predicted Mol.Wt.: (Calculated)52kDa.

Subcellular Location:

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.

Tissue Specificity:

Q15796 SMAD2_HUMAN:
Expressed at high levels in skeletal muscle, endothelial cells, heart and placenta.

Sequence:
MSSILPFTPPVVKRLLGWKKSAGGSGGAGGGEQNGQEEKWCEKAVKSLVKKLKKTGRLDELEKAITTQNCNTKCVTIPSTCSEIWGLSTPNTIDQWDTTGLYSFSEQTRSLDGRLQVSHRKGLPHVIYCRLWRWPDLHSHHELKAIENCEYAFNLKKDEVCVNPYHYQRVETPVLPPVLVPRHTEILTELPPLDDYTHSIPENTNFPAGIEPQSNYIPETPPPGYISEDGETSDQQLNQSMDTGSPAELSPTTLSPVNHSLDLQPVTYSEPAFWCSIAYYELNQRVGETFHASQPSLTVDGFTDPSNSERFCLGLLSNVNRNATVEMTRRHIGRGVRLYYIGGEVFAECLSDSAIFVQSPNCNQRYGWHPATVCKIPPGCNLKIFNNQEFAALLAQSVNQGFEAVYQLTRMCTIRMSFVKGWGAEYRRQTVTSTPCWIELHLNGPLQWLDKVLTQMGSPSVRCSSMS

Research Background

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 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.

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 (PubMed:21947082). 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.

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).

Extracellular region or secreted Cytosol Plasma membrane Cytoskeleton Lysosome Endosome Peroxisome ER Golgi apparatus Nucleus Mitochondrion Manual annotation Automatic computational assertionGraphics by Christian Stolte

Tissue Specificity:

Expressed at high levels in skeletal muscle, endothelial cells, heart and placenta.

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 PH domain). 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 (PubMed:24324267). Identified in a complex that contains at least ZNF451, SMAD2, SMAD3 and SMAD4 (PubMed:24324267). 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.

Research Fields

Research Fields:

· Cellular Processes > Cell growth and death > Cell cycle.(View pathway)
· Cellular Processes > Transport and catabolism > Endocytosis.(View pathway)
· Cellular Processes > Cellular community - eukaryotes > Adherens junction.(View pathway)
· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.(View pathway)
· Cellular Processes > Cell growth and death > Cellular senescence.(View pathway)
· Environmental Information Processing > Signal transduction > Hippo signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > FoxO signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > Apelin signaling pathway.(View pathway)
· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.(View pathway)
· Human Diseases > Cancers: Specific types > Pancreatic cancer.(View pathway)
· Human Diseases > Cancers: Overview > Pathways in cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Gastric cancer.(View pathway)
· Human Diseases > Cancers: Specific types > Colorectal cancer.(View pathway)
· Human Diseases > Cancers: Overview > Proteoglycans in cancer.
· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma.(View pathway)
· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).
· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).
· Human Diseases > Infectious diseases: Viral > HTLV-I infection.
· Organismal Systems > Endocrine system > Relaxin signaling pathway.
· Organismal Systems > Immune system > Th17 cell differentiation.(View pathway)

Reference Citations:

1). Wang XF;Fang QQ;Jia B;Hu YY;Wang ZC;Yan KP;Yin SY;Liu Z;Tan WQ; et al. Potential Effect of Non-Thermal Plasma for the Inhibition of Scar Formation: A Preliminary Report. Sci Rep 2020 Jan 23;10(1):1064. (PubMed: 31974451) [IF=4.011]

2). Liu S et al. Sphingomyelin synthase 1 regulates the epithelial‑to‑mesenchymal transition mediated by the TGF‑β/Smad pathway in MDA‑MB‑231 cells. Mol Med Rep 2018 Dec 4 (PubMed: 30535436)

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Catalog Number :

AF8314-BP
(Blocking peptide available as AF8314-BP)

Price/Size :

$350/1mg.
Tips: For phospho antibody, we provide phospho peptide(0.5mg) and non-phospho peptide(0.5mg).

Function :

Blocking peptides are peptides that bind specifically to the target antibody and block antibody binding. These peptide usually contains the epitope recognized by the antibody. Antibodies bound to the blocking peptide no longer bind to the epitope on the target protein. This mechanism is useful when non-specific binding is an issue, for example, in Western blotting (immunoblot) and immunohistochemistry (IHC). By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific; Specific binding will be absent from the western blot or immunostaining performed with the neutralized antibody.

Format and storage :

Synthetic peptide was lyophilized with 100% acetonitrile and is supplied as a powder. Reconstitute with 0.1 ml DI water for a final concentration of 10 mg/ml.The purity is >90%,tested by HPLC and MS.Storage Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C.

Precautions :

This product is for research use only. Not for use in diagnostic or therapeutic procedures.

Xenopus
100%
Zebrafish
100%
Rabbit
100%
Pig
100%
Dog
100%
Sheep
100%
Bovine
100%
Horse
0%
Chicken
0%
High similarity Medium similarity Low similarity No similarity
Q15796 as Substrate
Site PTM Type Enzyme
Ubiquitination
S2 Acetylation
S2 Phosphorylation
T8 Phosphorylation P27361 (MAPK3) , P24941 (CDK2)
K13 Ubiquitination
K19 Acetylation
K20 Acetylation
S21 Phosphorylation
K39 Acetylation
K46 Sumoylation
S47 Phosphorylation
K63 Ubiquitination
Y102 Phosphorylation
S110 Phosphorylation Q9UQM7 (CAMK2A) , Q9H4A3 (WNK1)
S118 Phosphorylation
K121 Ubiquitination
Y128 Phosphorylation
K156 Sumoylation
K156 Ubiquitination
K157 Ubiquitination
Y165 Phosphorylation
T172 Phosphorylation
T197 Phosphorylation P25098 (GRK2)
T220 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
S240 Phosphorylation Q9UQM7 (CAMK2A)
S245 Phosphorylation Q14680 (MELK) , P28482 (MAPK1) , P27361 (MAPK3)
S250 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
S255 Phosphorylation P28482 (MAPK1) , P27361 (MAPK3)
S260 Phosphorylation Q9UQM7 (CAMK2A) , Q9H4A3 (WNK1)
S317 Phosphorylation
T324 Phosphorylation
S417 Phosphorylation Q13177 (PAK2)
K420 Acetylation
S458 Phosphorylation
S460 Phosphorylation
S464 Phosphorylation P36897 (TGFBR1) , O00238 (BMPR1B)
S465 Phosphorylation Q9H4A3 (WNK1) , P36897 (TGFBR1) , O00238 (BMPR1B) , O96013 (PAK4) , Q8NER5 (ACVR1C)
S467 Phosphorylation O00238 (BMPR1B) , P36897 (TGFBR1) , Q8NER5 (ACVR1C)
IMPORTANT: For western blots, incubate membrane with diluted antibody in 5% w/v milk , 1X TBS, 0.1% Tween®20 at 4°C with gentle shaking, overnight.

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