Phospho-SMAD6 (Ser435) Antibody | Affinity Biosciences

Cites

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Expression of pSMAD1/5/8, pSMAD6, RUNX2 and pERK in knee articular cartilage of 9.

Product:	Phospho-SMAD6 (Ser435) Antibody
Catalog:	AF3769
Description:	Rabbit polyclonal antibody to Phospho-SMAD6 (Ser435)
Application:	ELISA(peptide)
Reactivity:	Human, Mouse, Rat
Mol.Wt.:	53kD(Calculated).
Uniprot:	O43541
RRID:	AB_2847083

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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. By comparing the staining from the blocked antibody versus the antibody alone, one can see which staining is specific.

Size	Price	Inventory
100ul	$350	In stock
200ul	$450	In stock

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Related Downloads

MSDS

Data Sheet

COA

Protocols

WB Handbook

IHC Protocol

IF/ICC Protocol

Product Info

Source:

Rabbit

Application:

ELISA(peptide) 1:20000-1:40000
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:

Human,Mouse,Rat

Clonality:

Polyclonal

Specificity:

Phospho-SMAD6 (Ser435) Antibody detects endogenous levels of SMAD6 only when phosphorylated at Ser435.

RRID:

AB_2847083
Cite Format: Affinity Biosciences Cat# AF3769, RRID:AB_2847083.

Conjugate:

Unconjugated.

Purification:

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

Storage:

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.

Alias:

Fold/Unfold

hSMAD6; MAD homolog 6; MADH6; MADH7; Mothers against decapentaplegic homolog 6; Mothers against DPP homolog 6; SMAD 6; SMAD family member 6; SMAD mothers against DPP homolog 6; Smad6; SMAD6_HUMAN;

Immunogens

Immunogen:

A synthesized peptide derived from human SMAD6 around the phosphorylation site of Ser435.

Uniprot:

O43541(SMAD6_HUMAN) >>Visit HPA database.

Gene(ID):

SMAD6(4091)

Expression:

O43541 SMAD6_HUMAN:

Ubiquitous in various organs, with higher levels in lung. Isoform B is up-regulated in diseased heart tissue.

Sequence:

O43541 SMAD6_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MFRSKRSGLVRRLWRSRVVPDREEGGSGGGGGGDEDGSLGSRAEPAPRAREGGGCGRSEVRPVAPRRPRDAVGQRGAQGAGRRRRAGGPPRPMSEPGAGAGSSLLDVAEPGGPGWLPESDCETVTCCLFSERDAAGAPRDASDPLAGAALEPAGGGRSREARSRLLLLEQELKTVTYSLLKRLKERSLDTLLEAVESRGGVPGGCVLVPRADLRLGGQPAPPQLLLGRLFRWPDLQHAVELKPLCGCHSFAAAADGPTVCCNPYHFSRLCGPESPPPPYSRLSPRDEYKPLDLSDSTLSYTETEATNSLITAPGEFSDASMSPDATKPSHWCSVAYWEHRTRVGRLYAVYDQAVSIFYDLPQGSGFCLGQLNLEQRSESVRRTRSKIGFGILLSKEPDGVWAYNRGEHPIFVNSPTLDAPGGRALVVRKVPPGYSIKVFDFERSGLQHAPEPDAADGPYDPNSVRISFAKGWGPCYSRQFITSCPCWLEILLNNPR

PTMs - O43541 As Substrate

O43541

Site	PTM Type	Enzyme	Source
R75	Methylation		Uniprot
K173	Ubiquitination		Uniprot
S274	Phosphorylation		Uniprot
S280	Phosphorylation		Uniprot
S283	Phosphorylation		Uniprot
S435	Phosphorylation	P51817 (PRKX)	Uniprot
K470	Ubiquitination		Uniprot

Research Backgrounds

O43541_SMAD6_HUMAN

Function:

Acts as a mediator of TGF-beta and BMP antiflammatory activity. Suppresses IL1R-TLR signaling through its direct interaction with PEL1, preventing NF-kappa-B activation, nuclear transport and NF-kappa-B-mediated expression of proinflammatory genes. May block the BMP-SMAD1 signaling pathway by competing with SMAD4 for receptor-activated SMAD1-binding. Binds to regulatory elements in target promoter regions.

PTMs:

Phosphorylated by BMP type 1 receptor kinase and by PRKX.

Monoubiquitinated at Lys-173 by the E2/E3 hybrid ubiquitin-protein ligase UBE2O, leading to reduced binding affinity for the activated BMP type I receptor ACVR1/ALK2, thereby enhancing BMP7 and regulating adipocyte differentiation. Ubiquitinated by WWP1 (By similarity). Ubiquitinated by RNF165, promoting proteasomal degradation, leading to enhance the BMP-Smad signaling (By similarity).

Arginine methylation by PRMT1, which is recruited by BMPR2, initiates BMP-Induced signaling and induces dissociation from the BMPR1B receptor at the cell surface leading to derepress downstream Smad1/Smad5 signaling.

Subcellular Location:

Nucleus.

Tissue Specificity:

Ubiquitous in various organs, with higher levels in lung. Isoform B is up-regulated in diseased heart tissue.

Subunit Structure:

Interacts with NEDD4L (By similarity). Interacts with WWP1 (By similarity). Interacts with STAMBP and PRKX. Interacts with RNF111 and AXIN1. Interacts with TGF-beta type I receptor superfamily members, including ACVR1B, BMPR1B and TGFBR1. In response to BMP2, but not to TGFB treatment, interacts with SMAD1, but not with SMAD2, nor with SMAD4; this interaction may inhibit SMAD1 binding to SMAD4. Interacts with HOXC8 and HOXC9. Interacts with PELI1; this interaction interferes with PELI1 complex formation with TRAF6, IRAK1, IRAK4 and MYD88 in response to IL1B and hence negatively regulates IL1R-TLR signaling.

Family&Domains:

Belongs to the dwarfin/SMAD family.

Research Fields

· Environmental Information Processing > Signal transduction > TGF-beta signaling pathway. (View pathway)

References

1). FGF receptor inhibitor BGJ398 partially rescues osteoarthritis-like phenotype in older high molecular weight FGF2 transgenic mice via multiple mechanisms. Scientific Reports (PubMed: 36153352) [IF=4.6]

Application: IHC Species: Mouse Sample:

Expression of pSMAD1/5/8, pSMAD6, RUNX2 and pERK in knee articular cartilage of 9.5-month-old Vector-Control and HMWTgFGF2 mice treated with BGJ398. Eight month-old Vector-Control (Vec) and HMWTgFGF2 (HMW) female mice were sq injected with vehicle or BGJ398 5 days/week for 6 weeks. At the end of the experiment (9.5 month old) knee joints were collected for IHC staining. Representative immunohistochemical staining images for (A) pSMAD1/5/8, (B) pSMAD6, (C) RUNX2, and (D) pERK. Quantitative analysis of percent-positive staining area for (E) pSMAD1/5/8, (F) pSMAD6, (G) RUNX2, and (H) pERK in articular cartilage. Data are shown as mean and individual points. n = 7–9 mice/group for (E, G, H); n = 4 mice /group for (F). *p

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Phospho-SMAD6 (Ser435) Antibody - #AF3769