Product Name	Phospho-Smad2 (Ser465 + Ser467)
Chinese Name	磷酸化细胞Signal transduction分子SMAD2抗体
Alias	Smad2(Phospho S465 + S467); phospho-Smad2(p-Ser465/467); p-Smad2(Ser465/467); phospho-Smad2(p-S465/467); Smad2 (phospho S465 + S467); p-Smad2 (phospho S465 + S467); hMAD 2; hSMAD2; JV18 1; JV18; JV181; MAD; MAD Related Protein 2; MADH2; MADR2; MGC22139; MGC34440; Mothers Against Decapentaplegic Homolog 2; mothers against DPP homolog 2; SMAD 2; SMAD; SMAD2; SMAD2_HUMAN.
literatures	Specific References  (12)     |     SL3419R has been referenced in 12 publications. [IF=13.751] Yvette Robbins. et al. Dual PD-L1 and TGF-b blockade in patients with recurrent respiratory papillomatosis. J Immunother Cancer. 2021 Aug;9(8):e003113  IF ;  Human.   PubMed:34462327 [IF=5.714] Han B et al. Deltamethrin induces liver fibrosis in quails via activation of the TGF-β1/Smad signaling pathway. Environ Pollut. 2019 Dec 23;259:113870.  WB ;  quail.   PubMed:31918140 [IF=5.589] Lv Y et al. Imidacloprid-induced liver fibrosis in quails via activation of the TGF-β1/Smad pathway. Sci Total Environ. 2019 Dec 6;705:135915.  WB ;  Quail.   PubMed:31835194 [IF=5.458] Chong Yin. et al. Long noncoding RNA Lnc-DIF inhibits bone formation by sequestering miR-489-3p. Iscience. 2022 Mar;25:103949  WB ;  Mouse.   PubMed:10.1016/j.isci.2022.103949 [IF=4.522] Ge Y et al. TCEA3 promotes differentiation of C2C12 cells via an Annexin A1‐mediated transforming growth factor‐β signaling pathway. J Cell Physiol. 2019 Jul;234(7):10554-10565.  WB&ICF ;  Mouse.   PubMed:30623413 [IF=3.69] Naihua Hu. et al. Forsythiae Fructuse water extract attenuates liver fibrosis via TLR4/MyD88/NF-κB and TGF-β/smads signaling pathways. J Ethnopharmacol. 2020 Nov;262:113275  WB ;  Rat.   PubMed:32810620 [IF=3.06] Yan Y et al. Inhibition of TGF-β Signaling in Gliomas by the Flavonoid Diosmetin Isolated from Dracocephalum peregrinum L. Molecules. 2020 Jan 2;25(1). pii: E192.  WB ;  Human.   PubMed:31906574 [IF=3.043] Yi X et al. Low‐intensity pulsed ultrasound protects subchondral bone in rabbit temporomandibular joint osteoarthritis by suppressing TGF‐β1/Smad3 pathway. J Orthop Res. 2020 Feb 15.  IHC-P ;  Rabbit.   PubMed:32060941 [IF=2.819] Lian L et al. Anti-fibrogenic Potential of Mesenchymal Stromal Cells in Treating Fibrosis in Crohn's Disease.Dig Dis Sci. 2018 Jul;63(7):1821-1834.  WB ;  Mouse.   PubMed:29704139 [IF=2.795] Chunyu Zhang et al. WISP1 promotes bovine MDSC differentiation via recruitment of ANXA1 for the regulation of the TGF-β signalling pathway. Mol Cell Biochem. 2020 Jul;470(1-2):215-227.  WB ;  Bovine.   PubMed:32458119 [IF=2.784] Zhang et al. Adenovirus‑mediated knockdown of activin A receptor type 2A attenuates immune‑induced hepatic fibrosis in mice and inhibits interleukin‑17‑induced activation of primary hepatic stellate cells. (2018) Int.J.Mol.Med. 42:279-289  WB ;  mouse.   PubMed:29620144 [IF=2.571] Tang et al. Salidroside protects against bleomycin-induced pulmonary fibrosis: activation of Nrf2-antioxidant signaling, and inhibition of NF-κB and TGF-β1/Smad-2/-3 pathways. (2016) Cell.Stress.Chaperones. 21:239-49  WB ;  Rat.   PubMed:26577463
Product Type	Phosphorylated anti
Research Area	immunology  Signal transduction  Apoptosis  transcriptional regulatory factor
Immunogen Species	Rabbit
Clonality	Polyclonal
React Species	Human, Mouse, Rat,  (predicted: Chicken, Dog, Cow, Horse, )
Applications	WB=1:500-2000 ELISA=1:5000-10000 IHC-P=1:100-500 IHC-F=1:100-500 Flow-Cyt=1ug/Test ICC=1:100 IF=1:100-500 （Paraffin sections need antigen repair） not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user.
Theoretical molecular weight	58kDa
Cellular localization	The nucleus cytoplasmic
Form	Liquid
Concentration	1mg/ml
immunogen	KLH conjugated Synthesised phosphopeptide derived from human SMAD2 around the phosphorylation site of Ser465/467: CS(p-S)M(p-S)
Lsotype	IgG
Purification	affinity purified by Protein A
Buffer Solution	0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
Storage	Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
Attention	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
PubMed	PubMed
Product Detail	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] 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. Subunit: Momomer; 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. Interacts with NEDD4L in response to TGF-beta. 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 and SKI. 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 ot the TGF-beta signaling. Interacts with PDPK1 (via PH domain). 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. On dephosphorylation by phosphatase PPM1A, released from the SMAD2/SMAD4 complex, and exported out of the nucleus by interaction with RANBP1. Tissue Specificity: Expressed at high levels in skeletal muscle, heart and placenta. 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. 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. 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. Similarity: Belongs to the dwarfin/SMAD family. Contains 1 MH1 (MAD homology 1) domain. Contains 1 MH2 (MAD homology 2) domain. SWISS: Q15796 Gene ID: 4087 Database links: Entrez Gene: 4087 Human Entrez Gene: 17126 Mouse Entrez Gene: 29357 Rat Omim: 601366 Human SwissProt: Q15796 Human SwissProt: Q62432 Mouse SwissProt: O70436 Rat Unigene: 12253 Human Unigene: 705764 Human Unigene: 391091 Mouse Unigene: 2755 Rat
Product Picture	Sample:Muscle (Mouse)Lysate at 40 ug Primary: Anti-p-Smad2(Ser465+Ser467)(SL3419R)at 1/300 dilution Secondary: IRDye800CW Goat Anti-RabbitIgG at 1/20000 dilution Predicted band size: 58kD Observed band size: 63kD Sample: Lane 1: Placenta (Mouse) Lysate at 40 ug Lane 2: Raw264.7 (Mouse) Cell Lysate at 30 ug Lane 3: Testis (Rat) Lysate at 40 ug Lane 4: Hela (Human) Cell Lysate at 30 ug Lane 5: HT1080 (Human) Cell Lysate at 30 ug Lane 6: Jurkat (Human) Cell Lysate at 30 ug Lane 7: HL60 (Human) Cell Lysate at 30 ug Primary: Anti-Phospho-Smad2 (Ser465 + Ser467) (SL3419R) at 1/1000 dilution Secondary: IRDye800CW Goat Anti-Rabbit IgG at 1/20000 dilution Predicted band size: 60 kD Observed band size: 60 kD Paraformaldehyde-fixed, paraffin embedded (Mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-Smad2(Ser465 + Ser467)) Polyclonal Antibody, Unconjugated (SL3419R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (Mouse testis); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (Phospho-Smad2(Ser465 + Ser467)) Polyclonal Antibody, Unconjugated (SL3419R) at 1:400 overnight at 4°C, followed by operating according to SP Kit(Rabbit) (sp-0023) instructionsand DAB staining. Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (Phospho-Smad2 (Ser465 + Ser467) ) polyclonal Antibody, Unconjugated (SL3419R) 1:100, 90 minutes at 37°C; followed by a conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei. Blank control（black line）:Hela. Primary Antibody (green line): Rabbit Anti-Phospho-Smad2 (Ser465 + Ser467) antibody (SL3419R) Dilution:1ug/Test; Secondary Antibody（white blue line）: Goat anti-rabbit IgG-AF488 Dilution: 0.5ug/Test. Isotype control（orange line）: Normal Rabbit IgG Protocol The cells were fixed with 4% PFA (10min at room temperature)and then permeabilized with 90% ice-cold methanol for 20 min at -20℃, The cells were then incubated in 5%BSA to block non-specific protein-protein interactions for 30 min at room temperature .Cells stained with Primary Antibody for 30 min at room temperature. The secondary antibody used for 40 min at room temperature. Acquisition of 20,000 events was performed.

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