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  • Product Name
    Phospho-Smad1/5/9 (Ser463+Ser465) Antibody
  • Catalog No.
    AF8313
  • RRID
    AB_2840375
  • Source
    Rabbit
  • Application
    WB,IHC,ELISA(peptide)
  • Reactivity
    Human, Rat, Monkey
  • Prediction
    Pig, Zebrafish, Bovine, Sheep, Rabbit, Dog, Chicken, Xenopus
  • UniProt
  • Mol.Wt
    56kD;
    52kDa(Calculated).
  • Concentration
    1mg/ml
  • Browse similar products>>

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Product Information

Alternative Names:Expand▼

BSP-1; BSP1; HsMAD1; JV4-1; JV41; MAD homolog 1; MAD mothers against decapentaplegic homolog 1; Mad related protein 1; Mad-related protein 1; MADH1; MADR1; Mothers against decapentaplegic homolog 1; Mothers against DPP homolog 1; SMA- AND MAD-RELATED PROTEIN 1; SMAD 1; SMAD family member 1; SMAD mothers against DPP homolog 1; Smad1; SMAD1_HUMAN; TGF beta signaling protein 1; Transforming growth factor-beta-signaling protein 1; DKFZp781C1895; DKFZp781O1323; Dwfc; hSmad5; JV5 1; JV5-1; MAD homolog 5; MAD, mothers against decapentaplegic homolog 5; MADH 5; MADH5; Mothers against decapentaplegic homolog 5; mothers against decapentaplegic, drosophila, homolog of, 5; Mothers against DPP homolog 5; MusMLP; SMA and MAD related protein 5; SMAD 5; SMAD family member 5; SMAD, mothers against DPP homolog 5; Smad5; SMAD5_HUMAN; MAD homolog 9; Madh6; Mothers against decapentaplegic; Mothers against decapentaplegic homolog 9; Mothers against DPP homolog 9; SMAD 9; SMAD family member 9; SMAD, mothers against DPP homolog 9 (Drosophila); SMAD8A; SMAD8B; Smad9; SMAD9_HUMAN;

Applications:

WB 1:1000-3000, IHC 1:50-1:200, ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.

Reactivity:

Human, Rat, Monkey

Predicted Reactivity:

Pig, Zebrafish, Bovine, Sheep, Rabbit, Dog, Chicken, Xenopus

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-Smad1/5/9 (Ser463+Ser465) Antibody detects endogenous levels of Smad1/5/9 only when phosphorylated at Ser463+Ser465.

RRID:

AB_2840375
Please cite this product as: Affinity Biosciences Cat# AF8313, RRID:AB_2840375.

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 in 3D

Immunogen:

A synthesized peptide derived from human Smad1/5/9 around the phosphorylation site of Ser463+Ser465.

Uniprot:



>>Visit The Human Protein Atlas

Gene ID:

Gene Name:

SMAD5,SMAD1,SMAD9

Molecular Weight:

Observed Mol.Wt.: 56kD.
Predicted Mol.Wt.: 52kDa(Calculated)..

Subcellular Location:

Cytoplasm. Nucleus. Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4. Co-localizes with LEMD3 at the nucleus inner membrane.

Tissue Specificity:

Q15797 SMAD1_HUMAN:
Ubiquitous. Highest expression seen in the heart and skeletal muscle.

Q99717 SMAD5_HUMAN:
Ubiquitous.

O15198 SMAD9_HUMAN:
Expressed in heart, brain, placenta, lung, skeletal muscle, prostate, testis, ovary and small intestine. Also expressed in fetal brain, lung and kidney.

Sequence:
MNVTSLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDALVKKLKKKKGAMEELEKALSCPGQPSNCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWPDLQSHHELKPLECCEFPFGSKQKEVCINPYHYKRVESPVLPPVLVPRHSEYNPQHSLLAQFRNLGQNEPHMPLNATFPDSFQQPNSHPFPHSPNSSYPNSPGSSSSTYPHSPTSSDPGSPFQMPADTPPPAYLPPEDPMTQDGSQPMDTNMMAPPLPSEINRGDVQAVAYEEPKHWCSIVYYELNNRVGEAFHASSTSVLVDGFTDPSNNKNRFCLGLLSNVNRNSTIENTRRHIGKGVHLYYVGGEVYAECLSDSSIFVQSRNCNYHHGFHPTTVCKIPSGCSLKIFNNQEFAQLLAQSVNHGFETVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKVLTQMGSPHNPISSVS

MTSMASLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDALVKKLKKKKGAMEELEKALSSPGQPSKCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWPDLQSHHELKPLDICEFPFGSKQKEVCINPYHYKRVESPVLPPVLVPRHNEFNPQHSLLVQFRNLSHNEPHMPQNATFPDSFHQPNNTPFPLSPNSPYPPSPASSTYPNSPASSGPGSPFQLPADTPPPAYMPPDDQMGQDNSQPMDTSNNMIPQIMPSISSRDVQPVAYEEPKHWCSIVYYELNNRVGEAFHASSTSVLVDGFTDPSNNKSRFCLGLLSNVNRNSTIENTRRHIGKGVHLYYVGGEVYAECLSDSSIFVQSRNCNFHHGFHPTTVCKIPSSCSLKIFNNQEFAQLLAQSVNHGFEAVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKVLTQMGSPLNPISSVS

MHSTTPISSLFSFTSPAVKRLLGWKQGDEEEKWAEKAVDSLVKKLKKKKGAMDELERALSCPGQPSKCVTIPRSLDGRLQVSHRKGLPHVIYCRVWRWPDLQSHHELKPLECCEFPFGSKQKEVCINPYHYRRVETPVLPPVLVPRHSEYNPQLSLLAKFRSASLHSEPLMPHNATYPDSFQQPPCSALPPSPSHAFSQSPCTASYPHSPGSPSEPESPYQHSVDTPPLPYHATEASETQSGQPVDATADRHVVLSIPNGDFRPVCYEEPQHWCSVAYYELNNRVGETFQASSRSVLIDGFTDPSNNRNRFCLGLLSNVNRNSTIENTRRHIGKGVHLYYVGGEVYAECVSDSSIFVQSRNCNYQHGFHPATVCKIPSGCSLKVFNNQLFAQLLAQSVHHGFEVVYELTKMCTIRMSFVKGWGAEYHRQDVTSTPCWIEIHLHGPLQWLDKVLTQMGSPHNPISSVS

Research Background

Function:

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD1 is a receptor-regulated SMAD (R-SMAD). SMAD1/OAZ1/PSMB4 complex mediates the degradation of the CREBBP/EP300 repressor SNIP1. May act synergistically with SMAD4 and YY1 in bone morphogenetic protein (BMP)-mediated cardiac-specific gene expression.

Post-translational Modifications:

Phosphorylation of the C-terminal SVS motif by BMP type 1 receptor kinase activates SMAD1 by promoting dissociation from the receptor and trimerization with SMAD4.

Ubiquitinated by SMAD-specific E3 ubiquitin ligase SMURF1, leading to its degradation. Monoubiquitinated, leading to prevent DNA-binding. Deubiquitination by USP15 alleviates inhibition and promotes activation of TGF-beta target genes. Dephosphorylation, probably by PPM1A, induces its export from the nucleus to the cytoplasm (By similarity).

Subcellular Location:

Cytoplasm. Nucleus.
Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4 (PubMed:15647271). Co-localizes with LEMD3 at the nucleus inner membrane (PubMed:15647271). Exported from the nucleus to the cytoplasm when dephosphorylated (By similarity).

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:

Ubiquitous. Highest expression seen in the heart and skeletal muscle.

Subunit Structure:

Found in a complex with SMAD4 and YY1. Interacts with HGS, NANOG and ZCCHC12 (By similarity). Upon C-terminus phosphorylation: forms trimers with another SMAD1 and the co-SMAD SMAD4. Interacts with PEBP2-alpha subunit, CREB-binding protein (CBP), p300, SMURF1, SMURF2, USP15 and HOXC8. Associates with ZNF423 or ZNF521 in response to BMP2 leading to activate transcription of BMP target genes. Interacts with SKOR1. Interacts (via MH2 domain) with LEMD3. Binding to LEMD3 results in at least a partial reduction of receptor-mediated phosphorylation. Forms a ternary complex with PSMB4 and OAZ1 before PSMB4 is incorporated into the 20S proteasome. Found in a macromolecular complex with FAM83G. Interacts (via MH2 domain) with FAM83G (via MH2 domain); in a SMAD4-independent manner. Interacts with ZC3H3 (By similarity). Interacts with TMEM119 (By similarity). Interacts (via MH1 and MH2 domains) with ZNF8 (By similarity). Interacts with RANBP3L; the interaction increases when SMAD1 is not phosphorylated and mediates SMAD1 nuclear export.

Similarity:

The MH2 domain mediates phosphorylation-dependent trimerization through L3 loop binding of phosphoserines in the adjacent subunit.

Belongs to the dwarfin/SMAD family.

Function:

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD5 is a receptor-regulated SMAD (R-SMAD).

Post-translational Modifications:

Phosphorylated on serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.

Ubiquitin-mediated proteolysis by SMAD-specific E3 ubiquitin ligase SMURF1.

Subcellular Location:

Cytoplasm. Nucleus.
Note: Cytoplasmic in the absence of ligand. Migrates to the nucleus when complexed with SMAD4.

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:

Ubiquitous.

Subunit Structure:

May form trimers with the co-SMAD SMAD4. Interacts with PEBP2-alpha subunit and SMURF1. Interacts with SUV39H1 and SUV39H2. Interacts (via MH2 domain) with LEMD3. Interacts with WWP1. Interacts with TMEM119 (By similarity). Interacts with ZNF8. Interacts with RANBP3L.

Similarity:

Belongs to the dwarfin/SMAD family.

Function:

Transcriptional modulator activated by BMP (bone morphogenetic proteins) type 1 receptor kinase. SMAD9 is a receptor-regulated SMAD (R-SMAD).

Post-translational Modifications:

Phosphorylated on serine by BMP (bone morphogenetic proteins) type 1 receptor kinase.

Subcellular Location:

Cytoplasm. Nucleus.
Note: In the cytoplasm in the absence of ligand. Migration to the nucleus when complexed with SMAD4 (By similarity).

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 in heart, brain, placenta, lung, skeletal muscle, prostate, testis, ovary and small intestine. Also expressed in fetal brain, lung and kidney.

Subunit Structure:

Interaction with the co-SMAD SMAD4. Interacts with PEBP2-alpha subunit. Interacts with RANBP3L.

Similarity:

Belongs to the dwarfin/SMAD family.

Research Fields

Research Fields:

· Cellular Processes > Cellular community - eukaryotes > Signaling pathways regulating pluripotency of stem cells.(View pathway)
· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway.(View pathway)

Reference Citations:

1). Kong Y et al. Magnesium-enriched microenvironment promotes odontogenic differentiation in human dental pulp stem cells by activating ERK/BMP2/Smads signaling. Stem Cell Res Ther 2019 Dec 10;10(1):378 (PubMed: 31823825) [IF=5.116]

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Catalog Number :

AF8313-BP
(Blocking peptide available as AF8313-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.

Pig
100%
Bovine
100%
Sheep
100%
Dog
100%
Xenopus
100%
Zebrafish
100%
Chicken
100%
Rabbit
100%
Horse
0%
High similarity Medium similarity Low similarity No similarity
Q15797/Q99717/O15198 as Substrate
Site PTM Type Enzyme
Ubiquitination
S11 Phosphorylation
K21 Ubiquitination
K32 Ubiquitination
S78 Phosphorylation
K81 Ubiquitination
Y88 Phosphorylation
K116 Sumoylation
K116 Ubiquitination
K118 Sumoylation
K118 Ubiquitination
S132 Phosphorylation
S144 Phosphorylation
Y146 Phosphorylation
S187 Phosphorylation P50750 (CDK9) , P49336 (CDK8) , P28482 (MAPK1)
S195 Phosphorylation P49336 (CDK8) , P50750 (CDK9) , P28482 (MAPK1)
T202 Phosphorylation P49841 (GSK3B)
S206 Phosphorylation P50750 (CDK9) , P28482 (MAPK1) , P49336 (CDK8) , P50613 (CDK7)
S210 Phosphorylation
S214 Phosphorylation P28482 (MAPK1) , P50750 (CDK9) , P49336 (CDK8)
S239 Phosphorylation Q13315 (ATM)
K269 Ubiquitination
S315 Phosphorylation
T322 Phosphorylation Q8N4C8 (MINK1) , Q9UKE5 (TNIK)
K418 Ubiquitination
S456 Phosphorylation
S462 Phosphorylation O00238 (BMPR1B)
S463 Phosphorylation Q05655 (PRKCD) , P36894 (BMPR1A) , O00238 (BMPR1B)
S465 Phosphorylation Q05655 (PRKCD) , O00238 (BMPR1B) , P36894 (BMPR1A)
Site PTM Type Enzyme
T2 Acetylation
S12 Phosphorylation
K22 Ubiquitination
K33 Ubiquitination
S57 Phosphorylation
S58 Phosphorylation
S63 Phosphorylation
K64 Ubiquitination
S79 Phosphorylation
K82 Ubiquitination
Y89 Phosphorylation
K117 Sumoylation
K119 Sumoylation
K119 Ubiquitination
Y128 Phosphorylation
S133 Phosphorylation
S152 Phosphorylation
S188 Phosphorylation
K306 Ubiquitination
S315 Phosphorylation
K418 Ubiquitination
S462 Phosphorylation
S463 Phosphorylation
S465 Phosphorylation
Site PTM Type Enzyme
K25 Ubiquitination
S40 Phosphorylation
S82 Phosphorylation
K85 Ubiquitination
Y92 Phosphorylation
T136 Phosphorylation Q16539 (MAPK14)
K159 Ubiquitination
S317 Phosphorylation
K420 Ubiquitination
S458 Phosphorylation
S464 Phosphorylation
S465 Phosphorylation
S467 Phosphorylation
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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