Phospho-Smad2 (Ser467) Antibody | Affinity Biosciences

Product:	Phospho-Smad2 (Ser467) Antibody
Catalog:	AF3449
Description:	Rabbit polyclonal antibody to Phospho-Smad2 (Ser467)
Application:	WB IHC IF/ICC
Reactivity:	Human, Mouse, Rat
Prediction:	Pig, Zebrafish, Bovine, Sheep, Rabbit, Dog, Xenopus
Mol.Wt.:	58kDa; 52kD(Calculated).
Uniprot:	Q15796
RRID:	AB_2834844

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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	$280	In stock
200ul	$350	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:

WB 1:500-1:2000, IHC 1:50-1:200, IF/ICC 1:100-1:500
*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

Prediction:

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

Clonality:

Polyclonal

Specificity:

Phospho-Smad2 (Ser467) Antibody detects endogenous levels of Smad2 only when phosphorylated at Serine 467.

RRID:

AB_2834844
Cite Format: Affinity Biosciences Cat# AF3449, RRID:AB_2834844.

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

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;

Immunogens

Immunogen:

Uniprot:

Q15796(SMAD2_HUMAN) >>Visit HPA database.

Gene(ID):

SMAD2(4087)

Expression:

Q15796 SMAD2_HUMAN:

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

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.

Sequence:

Q15796 SMAD2_HUMAN:
Protein BLAST With
NCBI/
ExPASy/
Uniprot

MSSILPFTPPVVKRLLGWKKSAGGSGGAGGGEQNGQEEKWCEKAVKSLVKKLKKTGRLDELEKAITTQNCNTKCVTIPSTCSEIWGLSTPNTIDQWDTTGLYSFSEQTRSLDGRLQVSHRKGLPHVIYCRLWRWPDLHSHHELKAIENCEYAFNLKKDEVCVNPYHYQRVETPVLPPVLVPRHTEILTELPPLDDYTHSIPENTNFPAGIEPQSNYIPETPPPGYISEDGETSDQQLNQSMDTGSPAELSPTTLSPVNHSLDLQPVTYSEPAFWCSIAYYELNQRVGETFHASQPSLTVDGFTDPSNSERFCLGLLSNVNRNATVEMTRRHIGRGVRLYYIGGEVFAECLSDSAIFVQSPNCNQRYGWHPATVCKIPPGCNLKIFNNQEFAALLAQSVNQGFEAVYQLTRMCTIRMSFVKGWGAEYRRQTVTSTPCWIELHLNGPLQWLDKVLTQMGSPSVRCSSMS

Predictions

Predictions:

Score>80(red) has high confidence and is suggested to be used for WB detection. *The prediction model is mainly based on the alignment of immunogen sequences, the results are for reference only, not as the basis of quality assurance.

Species

Results

Score

Pig

100

Bovine

100

Sheep

100

Dog

100

Xenopus

100

Zebrafish

100

Rabbit

100

Horse

Chicken

Model Confidence:
High(score>80) Medium(80>score>50) Low(score<50) No confidence

PTMs - Q15796 As Substrate

Q15796

Site	PTM Type	Enzyme	Source
	Ubiquitination		Uniprot
S2	Acetylation		Uniprot
S2	Phosphorylation		Uniprot
T8	Phosphorylation	P27361 (MAPK3) , P24941 (CDK2)	Uniprot
K13	Ubiquitination		Uniprot
K19	Acetylation		Uniprot
K20	Acetylation		Uniprot
S21	Phosphorylation		Uniprot
K39	Acetylation		Uniprot
K46	Sumoylation		Uniprot
S47	Phosphorylation		Uniprot
K63	Ubiquitination		Uniprot
Y102	Phosphorylation		Uniprot
S110	Phosphorylation	Q9UQM7 (CAMK2A) , Q9H4A3 (WNK1)	Uniprot
S118	Phosphorylation		Uniprot
K121	Ubiquitination		Uniprot
Y128	Phosphorylation		Uniprot
K156	Sumoylation		Uniprot
K156	Ubiquitination		Uniprot
K157	Ubiquitination		Uniprot
Y165	Phosphorylation		Uniprot
T172	Phosphorylation		Uniprot
T197	Phosphorylation	P25098 (GRK2)	Uniprot
T220	Phosphorylation	P28482 (MAPK1) , P27361 (MAPK3)	Uniprot
S240	Phosphorylation	Q9UQM7 (CAMK2A)	Uniprot
S245	Phosphorylation	Q14680 (MELK) , P28482 (MAPK1) , P27361 (MAPK3)	Uniprot
S250	Phosphorylation	P28482 (MAPK1) , P27361 (MAPK3)	Uniprot
S255	Phosphorylation	P28482 (MAPK1) , P27361 (MAPK3)	Uniprot
S260	Phosphorylation	Q9UQM7 (CAMK2A) , Q9H4A3 (WNK1)	Uniprot
S317	Phosphorylation		Uniprot
T324	Phosphorylation		Uniprot
S417	Phosphorylation	Q13177 (PAK2)	Uniprot
K420	Acetylation		Uniprot
S458	Phosphorylation		Uniprot
S460	Phosphorylation		Uniprot
S464	Phosphorylation	P36897 (TGFBR1) , O00238 (BMPR1B)	Uniprot
S465	Phosphorylation	Q9H4A3 (WNK1) , P36897 (TGFBR1) , O00238 (BMPR1B) , O96013 (PAK4) , Q8NER5 (ACVR1C)	Uniprot
S467	Phosphorylation	O00238 (BMPR1B) , P36897 (TGFBR1) , Q8NER5 (ACVR1C)	Uniprot

Research Backgrounds

Q15796_SMAD2_HUMAN

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.

PTMs:

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.

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).

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. 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).

Family&Domains:

Belongs to the dwarfin/SMAD family.

Research Fields

· Cellular Processes > Cell growth and death > Cell cycle. (View pathway)

· Cellular Processes > Transport and catabolism > Endocytosis. (View pathway)

· Cellular Processes > Cell growth and death > Cellular senescence. (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)

· Environmental Information Processing > Signal transduction > FoxO signaling pathway. (View pathway)

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

· Environmental Information Processing > Signal transduction > Apelin signaling pathway. (View pathway)

· Environmental Information Processing > Signal transduction > Hippo signaling pathway. (View pathway)

· Human Diseases > Infectious diseases: Parasitic > Chagas disease (American trypanosomiasis).

· Human Diseases > Infectious diseases: Viral > HTLV-I infection.

· Human Diseases > Cancers: Overview > Pathways in cancer. (View pathway)

· Human Diseases > Cancers: Overview > Proteoglycans in cancer.

· Human Diseases > Cancers: Specific types > Colorectal cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Pancreatic cancer. (View pathway)

· Human Diseases > Cancers: Specific types > Hepatocellular carcinoma. (View pathway)

· Human Diseases > Cancers: Specific types > Gastric cancer. (View pathway)

· Human Diseases > Immune diseases > Inflammatory bowel disease (IBD).

· Organismal Systems > Immune system > Th17 cell differentiation. (View pathway)

· Organismal Systems > Endocrine system > Relaxin signaling pathway.

References

1). Dietary protein levels changed the hardness of muscle by acting on muscle fiber growth and the metabolism of collagen in sub-adult grass carp (Ctenopharyngodon idella). Journal of Animal Science and Biotechnology, 2022 (PubMed: 36002862) [IF=7.0]

Application: WB Species: grass carp Sample: muscle

Fig. 3 Western blot analysis of relative protein expression in the muscle of grass carp. A Collagen transcription related factors; B Related factors regulating collagen mRNA translation; C Collagen degradation related factors. Values are means ± SD and n = 6 for each group. Different letters are significantly different (P < 0.05)

2). Timosaponin BII inhibits TGF‐β mediated epithelial‐mesenchymal transition through Smad‐dependent pathway during pulmonary fibrosis. PHYTOTHERAPY RESEARCH, 2023 (PubMed: 36807664) [IF=6.1]

3). Regorafenib-Attenuated, Bleomycin-Induced Pulmonary Fibrosis by Inhibiting the TGF-β1 Signaling Pathway. International Journal of Molecular Sciences, 2021 (PubMed: 33671452) [IF=5.6]

Application: WB Species: Mouse Sample: Mlg cells

Figure 3. Regorafenib down-regulates TGF-β1/Smad and TGF-β1/non-Smad signals in pulmonary fibroblasts. (A) CAGAmouse embryonic fibroblast (NIH-3T3) cells were exposed to TGF-β1 (5 ng/mL) or a series concentration (0–32 µM) in serum-free medium for 18 h; (B) Mlg cells were treated with TGF-β1 (5 ng/mL) and/or RG (2 µM, 4 µM) for 30 min, and Western blot was used to detect Smad3, Smad2, and their phosphorylation expression levels. Densitometric analysis are shown beside; (C) Mlg cells were incubated with RG (2 µM, 4 µM) and/or TGF-β1 (5 ng/mL) for 12 h to analyze the Erk1/2 and Akt and their phosphorylation levels by Western blotting. Densitometric analysis are shown beside; (D) BLM-PPF cells were incubated with RG (2 µM, 4 µM) for 12 h to analyze Erk1/2 and Akt and their phosphorylation levels by Western blotting. Densitometric analysis are shown beside. β-tubulin or GAPDH were used as a loading control. Data in (A–D) are means ± standard error of mean (SEM); ### p < 0.001, * p < 0.05, ** p < 0.01, *** p < 0.001 (one-way ANOVA), NS: not significant.

4). Does Chronic Pancreatitis in Growing Pigs Lead to Articular Cartilage Degradation and Alterations in Subchondral Bone?. International journal of molecular sciences, 2024 (PubMed: 38396667) [IF=5.6]

Application: IHC Species: pig Sample:

Figure 6 (a) Representative immunohistochemical (IHC) staining for TGF-β1, TGFβR1, TGFβR2, pSMAD2, and pSMAD4 in the articular cartilage of the distal femur from control (CONT) and chronic pancreatitis (CP) pigs. Scale bars represent 50 μm. (b) Bar graphs illustrating the optical density (OD) of expression for the analyzed proteins. Measurements were conducted on two histological samples per animal in at least ten randomly selected areas. Bars indicate mean values ± standard error (n = 5 pigs per group). Statistical significance is denoted as * p < 0.05; ** p < 0.01; *** p < 0.001.

5). Molecular mechanism of Gan-song Yin inhibiting the proliferation of renal tubular epithelial cells by regulating miR-21-5p in adipocyte exosomes. Journal of ethnopharmacology, 2024 (PubMed: 38043753) [IF=4.8]

Application: WB Species: Mouse Sample: TCMK-1 cells

Fig. 4. Mechanism of GSY on TCMK-1. A. Viability of TCMK-1 cells treated with different concentrations of GSY. B. TCMK-1 cell viability in each experimental group. C. Changes in TCMK-1 cell cycle level after GSY intervention. D. Changes in apoptosis level of TCMK-1 cells after GSY intervention. E. Expression of TGF-β1, SMAD2, SMAD3, SMAD7, p-SMAD2, p-SMAD3 and p-SMAD7 proteins in TCMK-1 cells after GSY intervention. F. Gene expression levels of TGF-β1, SMAD2, SMAD3 and SMAD7 in TCMK-1 cells after GSY intervention. *p < 0.05; **p < 0.01; ***p < 0.001. NC: Negative control; MOD: Model (60 mmol/L glucose). All experiments were repeated three times.

6). Enhancer of zeste homolog 2 modulates oxidative stress-mediated pyroptosis in vitro and in a mouse kidney ischemia-reperfusion injury model. FASEB JOURNAL, 2020 (PubMed: 31914694) [IF=4.4]

Application: WB Species: human Sample: HK-2 cells

FIGURE 7|EZH2 regulated Nox4 expression via the ALK5/Smad2/Smad3 pathway. D-G, HK-2 cells were transfected with an siRNA against EZH2 or a negative control siRNA (si-NC) for 48 h before being exposed to H/R. D-F, Western blot analysis for the protein expression of ALK5, Smad2, Smad3, p-Smad2, and Smad3 in the indicated groups and quantitative analysis of ALK5, p-Smad2, and p-Smad3, n = 3.

7). TFPI2 suppresses the interaction of TGF-β2 pathway regulators to promote endothelial-mesenchymal transition in diabetic nephropathy. Journal of Biological Chemistry, 2022 (PubMed: 35157852) [IF=4.0]

Application: WB Species: Human Sample: hRGECs

Figure 9 TFPI2 promotes TGF-β/Smad signaling activation. Human renal glomerular endothelial cells (hRGECs) were infected with adenovirus encoding shRNA targeting TFPI2 (TFPI2 shRNA) or overexpressing TFPI2 (TFPI2 OE), followed by stimulation of 5 ng/ml TGF-β2 for 48 h. A, the expression of SMAD7, TGFBR1, TGFBR2, SMAD2/3, and phospho-SMAD2/3 (p-SMAD2/3) was determined by Western blot. Semiquantitative analysis of (B) SMAD7, (C) TGFBR1, and (D) TGFBR2, as well as (E and F) the ratio of p-SMAD2/3 to SMAD2/3. G and H, immunofluorescent staining of SMAD2/3 in hRGECs. Yellow arrows indicated nuclear translocation of SMAD2/3. Data are shown as the mean ± SD (n = 3). ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001. TGF-β, transforming growth factor beta; TFP12, tissue factor pathway inhibitor 2.

8). Flavonoids of Coreopsis tinctoria Nutt alleviate the oxidative stress and inflammation of glomerular mesangial cells in diabetic nephropathy via RhoA/ROCK signaling. Journal of Functional Foods, 2022 [IF=3.8]

9). Inhibition of Endothelial-Mesenchymal Transition Mediated by Activin Receptor Type IIA Attenuates Valvular Injury Induced by Group A Streptococcus in Lewis Rats. Frontiers in bioscience (Landmark edition), 2025 (PubMed: 39862082) [IF=3.3]

10). RUNX3-dependent oxidative epithelial-to-mesenchymal transition in methamphetamine-induced chronic lung injury. Cell stress & chaperones, 2020 (PubMed: 32681471) [IF=3.3]

Application: WB Species: Rat Sample:

Fig. 5 RUNX3-related oxidative EMT induced by MA through TGF-β signaling. a, b The expression of RUNX3 after transfection by Western blot assay. c, d ROS levels in the alveolar epithelial cells by flow cytometry. The mean value in P1 is the average fluorescence intensity, as a statistical indicator, representing the expression of ROS. (e) TGF-β signaling expression by Western blot. f TGF-β 1 expression in alveolar epithelial cells in the various groups. g p-Smad2/Smad2 expression in alveolar epithelial cells in the various groups. h–k EMT marker proteins E-cadherin, ZO-1, and α-SMA expressed in alveolar epithelial cells in the various groups. l, m Immunofluorescence assay of α-SMA and ZO-1 in alveolar epithelial cells. After transfection for 72 h, the cells were treated with MA (5 mM, 48 h) and/or NAC (5 mM, 1 h), respectively. Data are presented as the mean ± SD. *P

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Phospho-Smad2 (Ser467) Antibody - #AF3449