SMAD3 (phospho-T179) antibody

• Catalog Number: orb345687
• Category: Antibodies
• Description: SMAD3 (phospho-T179) antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, WB
• Reactivity: Mouse
• Isotype: IgG
• Immunogen: Anti-SMAD3 pT179 antibody was prepared by repeated immunizations with a synthetic peptide corresponding to an internal region of human Smad3 protein surrounding amino acid residue 179.
• Concentration: 1.1 mg/mL
• Dilution range: ELISA: 1:15,000-1:75,000, WB: 1:1,000
• Form/Appearance: Liquid (sterile filtered)
• Purity: Anti-SMAD3 pT179 affinity-purified antibody is directed against the phosphorylated form of human Smad3 protein at the pT179 residue. The product was affinity purified from monospecific antiserum by immunoaffinity purification. Antiserum was first purified against the phosphorylated form of the immunizing peptide. The resultant affinity purified antibody was then cross adsorbed against the non-phosphorylated form of the immunizing peptide. Reactivity occurs against human Smad3 pT179 protein and the antibody is specific for the phosphorylated form of the protein. Reactivity with non-phosphorylated human Smad3 is minimal by ELISA and western blot. Expect reactivity against phosphorylated Smad2. Reactivity against other phosphorylated Smad family members is not known. A BLAST analysis was used to suggest cross reactivity with Smad3 from human, mouse, rat, pig, dog, and marmoset based on 100% sequence homology with the immunogen. Reactivity against homologues from other sources is not known.
• Conjugation: Unconjugated
• UniProt ID: P84022
• NCBI: NP_005893
• Storage: Store vial at -20° C or below prior to opening. This vial contains a relatively low volume of reagent (25 µL). To minimize loss of volume dilute 1:10 by adding 225 µL of the buffer stated above directly to the vial. Recap, mix thoroughly and briefly centrifuge to collect the volume at the bottom of the vial. Use this intermediate dilution when calculating final dilutions as recommended below. Store the vial at -20°C or below after dilution. Avoid cycles of freezing and thawing.
• Buffer/Preservatives: 0.01% (w/v) Sodium Azide
• Alternative names: rabbit anti-SMAD3 pT179 antibody, SMAD-3, SMAD 3,
• Note: For research use only
• Application notes: Anti-SMAD3 pT179 has been tested for use in ELISA and by western blot, and suitable by immunohistochemistry. Specific conditions for reactivity should be optimized by the end user. Expect a band approximately 48.1 kDa in size corresponding to human phosphorylated Smad3 protein by western blotting in the appropriate stimulated tissue or cell lysate or extract.
• Expiration Date: 12 months from date of receipt.

NMuMG mouse mammary epithelial cells were probed for the activation of Smad3 by detecting phosphorylation of threonine 179. The cells were either untreated or treated with TGF-beta, transferred to membranes and probed with Anti-SMAD3 pT179 (RABBIT) Antibody. The antibody detects only Smad3 in stimulated cells suggesting detection of phosphorylated SMAD3 at T179.

The SMAD pathway follows the canonical TGF-ß signaling pathway. TGF-ß dimers bind to a receptor thereby activating the pathway. The type I receptor then recruits and phosphorylates a receptor regulated SMAD (R-SMAD).i.e. SMAD2 or SMAD3. The R-SMAD then binds to the common SMAD (coSMAD) i.e. SMAD4, and forms a heterodimeric complex. This complex then enters the cell nucleus and acts as a transcription factor.