Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) Rekombinanter Antikörper

Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) Rekombinant Antikörper für WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Wirt / Isotyp

Kaninchen / IgG

Getestete Reaktivität

human und mehr (2)

Anwendung

WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR

Konjugation

Unkonjugiert

Publikationen(7)

CloneNo.

240826D11

Kat-Nr. : 80427-2-RR

Synonyme

P SMAD2, p SMAD2 (Ser465/467), P SMAD2 (Ser465/467)/SMAD3 (Ser423/425), P SMAD2 Ser465/467 /SMAD3 Ser423/425, p SMAD2/SMAD3

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  WB analysis using 80427-2-RR

  Non-treated and Calyculin A treated HEK-293 cells were subjected to SDS PAGE followed by western blot with 80427-2-RR (Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) antibody) at dilution of 1:2000 incubated at room temperature for 1.5 hours. The membrane was stripped and re-blotted with Alpha Tubulin (66031-1-Ig) antibody as a loading control.

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  IF Staining of HEK-293 using 80427-2-RR

  Immunofluorescent analysis of (4% PFA) fixed TGF beta 1 treated and untreated HEK-293 cells using Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) antibody (80427-2-RR, Clone: 240826D11 ) at dilution of 1:500 and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2).

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  FC experiment of HEK-293 using 80427-2-RR

  1X10^6 HEK-293 cells untreated (dashed lines) or treated with Calyculin A were intracellularly stained with 0.13 ug Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) Recombinant antibody (80427-2-RR, Clone:240826D11) and CoraLite®488-Conjugated Goat Anti-Rabbit IgG(H+L) (SA00013-2)(red), or 0.13 ug Rabbit IgG Isotype Control Recombinant Antibody (98136-1-RR, Clone: 240953C9) (blue). Cells were fixed with 4% PFA and permeabilized with 90% MeOH.

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  ChIP experiment of HaCaT using 80427-2-RR

  Chromatin was prepared from HaCaT cells treated with TGF-β1 (7 ng/ml) for 1 h. Cells were fixed with formaldehyde for 10 minutes. The ChIP was performed with 20 µg of cross-linked chromatin, 5 µg of Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) (80427-2-RR) or 5 ug of Normal Rabbit IgG (98136-1-RR), and 20 µl of Protein A Magarose Beads. The immunoprecipitated DNA was quantified by real-time PCR.

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Geprüfte Anwendungen

• Erfolgreiche Detektion in WB: Mit Calyculin A behandelte HEK-293-Zellen
• Erfolgreiche Detektion in IF/ICC: TGF beta 1 treated HEK-293 cells
• Erfolgreiche Detektion in FC (Intra): Mit Calyculin A behandelte HEK-293-Zellen
• Erfolgreiche Detektion in ChIP-qPCR: TGF-β1 (7 ng/ml, 1 h) HaCaT cells

Empfohlene Verdünnung

• Western Blot (WB): WB : 1:1000-1:4000
• Immunfluoreszenz (IF)/ICC: IF/ICC : 1:250-1:1000
• Durchflusszytometrie (FC) (INTRA): FC (INTRA) : 0.13 ug per 10^6 cells in a 100 µl suspension
• CHImmunpräzipitation (IP)-QPCR: CHIP-QPCR : 1:10-1:100

It is recommended that this reagent should be titrated in each testing system to obtain optimal results.

Sample-dependent, check data in validation data gallery

Veröffentlichte Anwendungen

• WB: See 7 publications below

Produktinformation

80427-2-RR bindet in WB, IF/ICC, FC (Intra), ELISA, ChIP-qPCR Phospho-SMAD2 (Ser465/467)/SMAD3 (Ser423/425) und zeigt Reaktivität mit human

• Getestete Reaktivität: human
• In Publikationen genannte Reaktivität: human, Maus, Ratte
• Wirt / Isotyp: Kaninchen / IgG
• Klonalität: Rekombinant
• Typ: Antikörper
• Immunogen: Peptid
• Vollständiger Name: SMAD family member 2
• Berechnetes Molekulargewicht: 467 aa, 52 kDa
• Beobachtetes Molekulargewicht: 60 kDa
• GenBank-Zugangsnummer: BC014840
• Gene symbol: SMAD2
• Gene ID (NCBI): 4087
• Konjugation: Unkonjugiert
• Form: Liquid
• Reinigungsmethode: Protein A purfication
• Lagerungspuffer: PBS with 0.02% sodium azide and 50% glycerol
• Lagerungsbedingungen: Bei -20°C lagern. Nach dem Versand ein Jahr lang stabil Aliquotieren ist bei -20^oC Lagerung nicht notwendig. 20ul Größen enthalten 0,1% BSA.

Hintergrundinformationen

SMAD2, also named as MADH2 and MADR2, belongs to the dwarfin/SMAD family, contains 1 MH1 (MAD homology 1) domain and 1 MH2 (MAD homology 2) domain. SMAD2 is a receptor-regulated SMAD(R-SMAD) that is an intracellular signal transducer and transcriptional modulator activated by TGF-beta and activin type 1 receptor kinases. This protein may act as a tumor suppressor in colorectal carcinoma. It is phosphorylated on one or several of Thr-220, Ser-245, Ser-250, and Ser-255. In response to TGF-beta, It is phosphorylated on Ser-465/467 by TGF-beta and activin type 1 receptor kinases, and then able to interact with SMURF2, recruiting other proteins, such as SNON, for degradation. In response to decorin, the naturally occurring inhibitor of TGF-beta signaling, it is phosphorylated on Ser-240 by CaMK2. It is phosphorylated by MAPK3 upon EGF stimulation; which increases transcriptional activity and stability, and is blocked by calmodulin. In response to TGF-beta, it is ubiquitinated by NEDD4L, which promotes its degradation. In response to TGF-beta signaling, it is acetylated on Lys-19 by coactivators, which increases transcriptional activity. The molecular weight of unphosphorylated forms of Smad2 is 52 kDa and phosphorylated forms of Smad2 is 58 kDa. (PMID: 9006934)

Publikationen

Species	Application	Title
mouse	WB	Phytomedicine; ErTao decoction alleviates liver fibrosis by suppressing STING-mediated macrophages and NLRP3 inflammasome activation; Authors - Hui Jia
rat	WB	Bioeng Transl Med; Reduced glutathione enhances adipose tissue-derived mesenchymal stem cell engraftment efficiency for liver fibrosis by targeting TGFβ1/SMAD3/NOX4 pathway; Authors - Shaoxiong Yu
human	WB	Sci Rep; Autophagy inhibition enhances antifibrotic potential of placental mesenchymal stem cells of fetal origin via regulating TGF-β1 mediated protein degradation of HGF; Authors - Wei Zhou
mouse	WB	Eur J Pharmacol; Omentin-1 attenuates atrial fibrillation via Src/PI3K/Akt signaling-mediated anti-fibrotic effects in cardiac fibroblasts; Authors - Tian-Lun Li
human	WB	Clin Proteomics; Multi-omics analyses, cell experiments, and network pharmacology tools identified key proteins and candidate drugs for alopecia areata treatment; Authors - Lingbo Bi
human	WB	Cell Signal; CRABP2 promotes peritoneal metastasis in CRC through TGF-β/Smad-mediated EMT signaling and invadopodia formation; Authors - Wende Hao