SMAD2 / SMAD3 (pSer423), (pSer465) Antikörper (PE)

Produktnummer ABIN1177178

Produktdetails

Target: SMAD2 / SMAD3

Bindungsspezifität: pSer423, pSer465

Reaktivität: Human, Maus, Ratte

Wirt: Maus

Klonalität: Monoklonal

Konjugat: PE

Applikation: Intracellular Staining (ICS)

Marke: BD Phosflow™

Aufreinigung: The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.

Immunogen: Phosphorylated Human Smad2 Peptide

Klon: O72-670

Isotyp: IgG1 kappa

Anwendungsinformationen

Applikationshinweise: This antibody conjugate is suitable for intracellular staining of human lymphoid cell lines, peripheral blood mononuclear cells, and mouse splenocytes using BD Cytofix™ Fixation Buffer or BD Phosflow™ Lyse/Fix Buffer with BD Phosflow™ Perm Buffer III. Prior to stimulation, cells were serum starved overnight at a density of 2-10 million cells/mL in flat-bottom 96- or 6-well tissue culture plates or in loosely capped, round-bottom tubes containing approximately 100 mL of cells in suspension.
Note:
1. Serum starvation for 2 hours following PBMC isolation was not sufficient to reduce basal phosphorylation of Smad2 and Smad3.
2. Do not mix or agitate untreated cells until just before the cells are ready to be fixed, since agitation of serum-starved mouse or human primary leukocytes prior to fixation increased Smad2/3 phosphorylation, even in the absence of exogenous TGF-beta.

Probenmenge: 5 μL

Beschränkungen: Nur für Forschungszwecke einsetzbar

Handhabung

Format: Liquid

Buffer: Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.

Konservierungsmittel: Sodium azide

Vorsichtsmaßnahmen: This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.

Lagerung: 4 °C

Informationen zur Lagerung: Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.

Referenzen

Martinez, Zhang, Reynolds, Tanaka, Chung, Liu, Robertson, Lin, Feng, Dong: "Smad2 positively regulates the generation of Th17 cells." in: The Journal of biological chemistry, Vol. 285, Issue 38, pp. 29039-43, (2010) (PubMed).

Shinto, Yashiro, Toyokawa, Nishii, Kaizaki, Matsuzaki, Noda, Kubo, Tanaka, Doi, Ohira, Muguruma, Sawada, Hirakawa: "Phosphorylated smad2 in advanced stage gastric carcinoma." in: BMC cancer, Vol. 10, pp. 652, (2010) (PubMed).

Rosendahl, Speletas, Leandersson, Ivars, Sideras: "Transforming growth factor-beta- and Activin-Smad signaling pathways are activated at distinct maturation stages of the thymopoeisis." in: International immunology, Vol. 15, Issue 12, pp. 1401-14, (2003) (PubMed).

Moustakas, Souchelnytskyi, Heldin: "Smad regulation in TGF-beta signal transduction." in: Journal of cell science, Vol. 114, Issue Pt 24, pp. 4359-69, (2002) (PubMed).

Abdollah, Macías-Silva, Tsukazaki, Hayashi, Attisano, Wrana: "TbetaRI phosphorylation of Smad2 on Ser465 and Ser467 is required for Smad2-Smad4 complex formation and signaling." in: The Journal of biological chemistry, Vol. 272, Issue 44, pp. 27678-85, (1997) (PubMed).

Heldin, Miyazono, ten Dijke: "TGF-beta signalling from cell membrane to nucleus through SMAD proteins." in: Nature, Vol. 390, Issue 6659, pp. 465-71, (1997) (PubMed).

Souchelnytskyi, Tamaki, Engström, Wernstedt, ten Dijke, Heldin: "Phosphorylation of Ser465 and Ser467 in the C terminus of Smad2 mediates interaction with Smad4 and is required for transforming growth factor-beta signaling." in: The Journal of biological chemistry, Vol. 272, Issue 44, pp. 28107-15, (1997) (PubMed).

Antigendetails

Target: SMAD2 / SMAD3

Hintergrund: The O72-670 monoclonal antibody specifically binds to the Smad2 protein phosphorylated at the Ser465/467 sites and the Smad3 protein phosphorylated at the Ser423/425 sites. Smad2 and Smad3 are members of the Smad superfamily with observed molecular weights of 60 kDa and 52 kDa, respectively. The Smad family consists of three subfamilies: receptor regulated Smads or R-Smads, including Smads1, 2, 3, 5, and 8, common partner Smad, or Co-Smad, including Smad4, and inhibitory Smads, or I-Smad, including Smads 6 and 7. Activation of TGF-beta superfamily serine/threonine kinase receptors, such as TGF-beta, activin and BMP receptors, by their bound ligands leads to the phosphorylation of R-Smads at several sites. It has been shown that the ligand-bound TGF-beta type I receptor directly phosphorylates Ser465 and Ser467 of Smad2 and Ser423 and Ser425 of Smad3. Phosphorylated R-Smads form complexes with Co-Smad and translocate into the nucleus to regulate transcription affecting a wide range of critical cellular processes including cell-fate determination, proliferation, morphogenesis, differentiation and apoptosis. The inhibitory Smads inhibit this pathway through two potential mechanisms: either by preventing R-Smads from binding to their corresponding receptors and/or by competing with Smad4, the Co-Smad, from binding to R-Smads. High level expression of phosphorylated Smad2 has been associated with poor prognosis in late stage gastric carcinoma. Roles for Smad2 have been described in thymopoiesis and the TGF-beta-mediated induction of regulatory T cells and Th17 cells. The specificity of the O72-670 mAb was confirmed by Western blot and immunohistochemistry using unconjugated antibody.
Synonyms: SMAD2, SMAD3, MADH2, MADH3, MAD homolog 2, MAD homolog 3