Phospho-SMAD

Western blot analysis of lysates from HeLa cells

Western blot analysis of lysates from C6 cells

Western blot using affinity purified anti-Smad3 pS423 pS425 antibody shows detection of endogenous Smad3 in stimulated cell lysates. Lysates were prepared from control cells (- lanes), or cells stimulated with 2 ng/ml TGF (+lanes) for 1 hour. This reagent recognizes phosphorylated Smad3 and has negligible reactivity against non-phosphorylated Smad3 protein. Personal Communication. Ying Zhang, NIH, CCR, Bethesda, MD.

Western blot analysis of various lysates using (TA381749) at 1:1000 dilution. Hela cells were treated by TGF-β (10 ng/ml) at 37℃ for 30 minutes.Secondary antibody: HRP-conjugated Goat anti-Rabbit IgG (H+L) at 1:10000 dilution.Lysates/proteins: 25μg per lane.Blocking buffer: 3% nonfat dry milk in TBST.Detection: ECL Basic Kit .Exposure time: 90s.

Western blot analysis of Phospho-Smad2 (Ser255) in K562 lysates using Phospho-Smad2 (Ser255) antibody.

Immunohistochemical analysis of paraffin-embedded human pancreas, using Phospho-Smad5 (S463/465) Antibody.

Immunohistochemical analysis of paraffin-embedded human transitional cell carcinoma of bladder, using Phospho-Smad2 (S255) Antibody.

Western blot analysis of Phospho-Smad3 (Ser425) in various lysates using Phospho-Smad3 (Ser425) antibody.

Western blot analysis of Phospho-Smad2/3 (Thr8) in various lysates using Phospho-Smad2/3 (Thr8) antibody.

Immunohistochemistry analysis of paraffin-embedded Human Skin Tissue using Phospho-Smad3 (Ser425) (7H6) antibody.High-pressure and temperature Sodium Citrate pH 6.0 was used for antigen retrieval.

Western blot (WB) analysis of p-Smad1 (S465) pAb at 1:500 dilution Lane1:A549 whole cell lysate(40ug) Lane2:A549 treated with UV for 5 minutes then repair for 1 hour whole cell lysate(40ug) Lane3:A549 treated with UV for 5 minutes then repair for 3 hours whole cell lysate(40ug) Lane4:A549 treated with UV for 5 minutes then repair for 6 hours whole cell lysate(40ug) Lane5:A549 treated with UV for 5 minutes then repair for 24 hours whole cell lysate(40ug) Lane6:The Uterus tissue lysate of Mouse(40ug) Lane7:The Uterus tissue lysate of Rat(40ug)

Western blot analysis of various lysates

Western blot analysis of lysates from NIH/3T3 cells

Western blot analysis of Phospho-Smad2 (Ser250) in mouse brain, mouse lung lysates using Phospho-Smad2 (Ser250) antibody.

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.

Western blot analysis of lysates from HeLa cells

Western Blot of Rabbit anti-Smad2 pS465pS467 antibody. Lane 1: MDA-MB-231 cells. Lane 2: MDA-MB-231 cells treated with TGF-β1 for 1h. Load: 20 µg per lane. Primary antibody: Smad2pS465pS467 antibody at 1:1000 for overnight at 4°C. Secondary antibody: IRDye800™ rabbit secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO/TBST overnight at 4°C. Predicted/Observed size: 52.3 kDa for Smad2pS465pS467. Other band(s): ~85kDa non-specific band.

Western blot analysis of various lysates

Western blot analysis of various lysates

Western blot analysis of Phospho-Smad5 (S463/S465) expression in (1) HeLa cell lysate; (2) HeLa cell treated with BMP-4 lysate.

Western blot analysis of Phospho-Smad1 (Ser463/Ser465) in A549, U2OS, C6 lysates using Phospho-Smad1 (Ser463/Ser465) antibody.

Western blot analysis of Phospho-Smad3 (Ser423/425) in Jurkat, C6, CHO-K1, Hela lysates using Phospho-Smad3 (Ser423/425) antibody.

Western blot (WB) analysis of Smad2/3 (phospho-T8) polyclonal antibody at 1:500 dilution Lane1:HEK293T whole cell lysate Lane2:HEK293T treated with TNFα(20ng/ml)for 10 min whole cell lysate Lane3:The Heart tissue lysate of Mouse

Western blot (WB) analysis of p-Smad3 (S204) pAb at 1:500 dilution Lane1:CT26 whole cell lysate(40ug) Lane2:C6 whole cell lysate(40ug) Lane3:H1792 whole cell lysate(40ug) Lane4:A375 whole cell lysate(40ug) Lane5:Hela whole cell lysate(40ug)

Western blot (WB) analysis of p-Smad3 (S213) pAb at 1:500 dilution Lane1:Hela whole cell lysate(40ug) Lane2:HEK293T whole cell lysate(40ug) Lane3:Panc1 whole cell lysate(20ug) Lane4:LOVO whole cell lysate(40ug) Lane5:C6 whole cell lysate(40ug) Lane6:AML-12 whole cell lysate(40ug)

Western blot (WB) analysis of p-Smad2/3 (T8) pAb at 1:500 dilution Lane1:HEK293T whole cell lysate(40µg) Lane2:Hela whole cell lysate(40µg) Lane3:The Lung tissue lysate of Mouse(40µg) Lane4:SGC7901 whole cell lysate(40µg) Lane5:H9C2 whole cell lysate(40µg)