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P smad3 9520

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P-Smad3 (9520) is a rabbit monoclonal antibody that detects endogenous levels of Smad3 protein only when phosphorylated at serine 423 and serine 425. This antibody is designed for use in western blotting applications.

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Lab products found in correlation

Smad3 9523, by Cell Signaling Technology (3 mentions) Erk1 2 9102, by Cell Signaling Technology (2 mentions) E cadherin, by BD (2 mentions) P smad2 3108, by Cell Signaling Technology (2 mentions) Smad2 3103, by Cell Signaling Technology (2 mentions) N cadherin, by BD (2 mentions) β actin, by Merck Group (2 mentions) Akt 4691, by Cell Signaling Technology (2 mentions) Anti mouse 5470 and anti rabbit 5151 secondary antibodies, by Cell Signaling Technology (2 mentions) P38 9212, by Cell Signaling Technology (2 mentions) Slug 9585, by Cell Signaling Technology (2 mentions) P akt 4058, by Cell Signaling Technology (2 mentions) Perk1 2 9101, by Cell Signaling Technology (2 mentions) Ros assay kit, by Beyotime (1 mentions) Mda assay kit, by Beyotime (1 mentions) Cat assay kit, by Beyotime (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Bicinchoninic acid bca protein assay kit, by Beyotime (1 mentions) Ly2109761 s2704, by Selleck Chemicals (1 mentions) Cell counting kit 8 cck 8, by MultiSciences Biotech (1 mentions)

Close spelling variants of this product

Smad3 (242 citations) P-Smad3 (240 citations)

8 protocols using p smad3 9520

1

Cell Viability and Oxidative Stress Assays

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Cell Counting Kit-8 (CCK-8) was purchased from MultiSciences Biotech Co. Ltd. (Hangzhou, China). RPMI 1640 medium was purchased from Jenom Biotech Co. Ltd. (Hangzhou, China). Fetal bovine serum (FBS) was purchased from Gibco® (Waltham, USA). Bicinchoninic acid (BCA) protein assay kit, ROS assay kit, CAT assay kit, and MDA assay kit were purchased from Beyotime Biotech Co. Ltd. (Suzhou, China). Antibodies GAPDH (ab181602), β-actin (ab8227), Nrf2 (ab137550), TGF-β1 (ab92486), COL1A1 (ab21286), COL3A1 (ab7778), 4-HNE (ab46545), 8-OHdG (ab62623), and GPx1 (ab22604) were obtained from Abcam plc. (Cambridge, UK). Antibodies elastin (sc58756), TIMP-1 (sc5538), TIMP-2 (sc5539), MMP-2 (sc10736), and MMP-9 (sc10737) were purchased from Santa Cruz Biotech Inc. (Dallas, USA). mTGF-β1 (5231LC) and antibodies Smad2/3 (8685), p-Smad2 (3108), and p-Smad3 (9520) were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Antibody MnSOD (06-984) was purchased from Millipore (Billerica, USA); LY2109761 (S2704) was purchased from Selleck (Houston, USA).
Tang J., Li B., Liu C., Li Y., Li Q., Wang L., Min J., Hu M., Hong S, & Hong L. (2017). Mechanism of Mechanical Trauma-Induced Extracellular Matrix Remodeling of Fibroblasts in Association with Nrf2/ARE Signaling Suppression Mediating TGF-β1/Smad3 Signaling Inhibition. Oxidative Medicine and Cellular Longevity, 2017, 8524353.
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2

Epithelial-Mesenchymal Transition Antibody Assay

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Antibodies against Slug (SC‐10436), Twist (SC‐15393) and β‐actin (sc‐47778) were purchased from Santa Cruz (CA, USA); those against cleaved caspase‐3 (9661), cleaved PARP (9541), TGF‐β (3711), Smad2 (5339), p‐Smad2 (3108), Smad3 (9523) and p‐Smad3 (9520) were from Cell Signaling Technology (MA, USA); and those against N‐cadherin (610921) and E‐cadherin (610182) were from BD, NJ, USA. Anti–HAS2 (ab140671), anti–Vimentin (3634‐100) and anti–zeb1 (HPA027524) were from Abcam (MA, USA), Biovision (CA, USA) and Sigma (MO, USA), respectively. Hyaluronan (GLR001, low molecular weight; GLR004, middle molecular weight; GLR002, high molecular weight) was purchased from R&D Systems (MN, USA). 4‐Methylumbelliferone (M1381) and SB431542 (S4317) was purchased from Sigma (MO, USA).
Kim Y., Lee S.B., Shim S., Kim A., Park J., Jang W., Lee S., Myung J.K., Park S., Lee S, & Kim M. (2019). Hyaluronic acid synthase 2 promotes malignant phenotypes of colorectal cancer cells through transforming growth factor beta signaling. Cancer Science, 110(7), 2226-2236.
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3

TGF-β and Copper Sulfate Regulation

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TGF-β (T7039) and copper sulfate (C8027) were obtained from Sigma-Aldrich (St. Louis, MO, USA). D-penicillamine (Sigma) and trientine hydrochloride (Sigma) were used as chelators of copper. Antibodies specific for β-actin (47778), Vimentin (6260), N-cadherin (59987), and E-cadherin (8426) were purchased from Santa Cruz Biotechnology. Antibodies for TGF-β (3711), Smad2/3 (3102), p-Smad2 (18338), and p-Smad3 (9520) were purchased from Cell Signaling Technology (Beverly, MA, USA).
Kim H., Jo S., Kim I.G., Kim R.K., Kahm Y.J., Jung S.H, & Lee J.H. (2022). Effect of Copper Chelators via the TGF-β Signaling Pathway on Glioblastoma Cell Invasion. Molecules, 27(24), 8851.
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4

TGF-β Signaling Pathway Activation

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Cells were serum-starved overnight and treated with 100pM TGF-β for 60 minutes. Cells were lysed in 2× Laemmli sample buffer, and the proteins were resolved on a 10% SDS-PAGE gel. All primary antibodies (Slug #9585, p-Smad2 #3108, Smad2 #3103, p-Smad3 #9520, Smad3 #9523, p-ERK1/2 #9101, ERK1/2 #9102, p-AKT #4058, AKT #4691, p-p38 #4631, p38 #9212) were purchased from Cell Signaling (Danvers, MA, USA) except for β-actin (Sigma, AA5441) and N-cadherin (#610921) and E-cadherin (#610182) (BD Biosciences). Anti-mouse (#5470) and anti-rabbit (#5151) secondary antibodies were purchased from Cell Signaling. Blots were scanned using the LI-COR Odyssey (LI-COR Biosciences, Lincoln, NE, USA).
Huang J.J., Corona A.L., Dunn B.P., Cai E.M., Prakken J.N, & Blobe G.C. (2019). Increased type III TGF-β receptor shedding decreases tumorigenesis through induction of epithelial-to-mesenchymal transition. Oncogene, 38(18), 3402-3414.
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5

TGF-β Signaling Pathway Activation

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Cells were serum-starved overnight and treated with 100pM TGF-β for 60 minutes. Cells were lysed in 2× Laemmli sample buffer, and the proteins were resolved on a 10% SDS-PAGE gel. All primary antibodies (Slug #9585, p-Smad2 #3108, Smad2 #3103, p-Smad3 #9520, Smad3 #9523, p-ERK1/2 #9101, ERK1/2 #9102, p-AKT #4058, AKT #4691, p-p38 #4631, p38 #9212) were purchased from Cell Signaling (Danvers, MA, USA) except for β-actin (Sigma, AA5441) and N-cadherin (#610921) and E-cadherin (#610182) (BD Biosciences). Anti-mouse (#5470) and anti-rabbit (#5151) secondary antibodies were purchased from Cell Signaling. Blots were scanned using the LI-COR Odyssey (LI-COR Biosciences, Lincoln, NE, USA).
Huang J.J., Corona A.L., Dunn B.P., Cai E.M., Prakken J.N, & Blobe G.C. (2019). Increased type III TGF-β receptor shedding decreases tumorigenesis through induction of epithelial-to-mesenchymal transition. Oncogene, 38(18), 3402-3414.
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6

Synthesis and Characterization of Novel Gold(I) Complex

Cited 1 time
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[Triphenylphosphine-1,3-diethylbenzylimidazol-2-ylidene)]gold(I) iodide (MC4) was synthesized as described19 (link),20 (link),52 (link). TCDD was purchased from Sigma Aldrich (Germany). Resveratrol and Auranofin were from Santa Cruz (Germany). Antibodies of AHR (Biomol, Germany, AP5533C, 1:1000) and CYP1A1 were obtained from Abgent (Biomol, Germany, F50940, 1:1000). ACTIN (SC-47778, 1:1000) and VINCULIN (SC-73614) were from Santa Cruz. SMAD4 (9515, 1:1000), pSMAD3 (9520), and SMAD3 (9523, 1:1000) were from Cell Signaling (NEB, Germany).
Cheng X., Haeberle S., Shytaj I.L., Gama-Brambila R.A., Theobald J., Ghafoory S., Wölker J., Basu U., Schmidt C., Timm A., Taškova K., Bauer A.S., Hoheisel J., Tsopoulidis N., Fackler O.T., Savarino A., Andrade-Navarro M.A., Ott I., Lusic M., Hadaschik E.N, & Wölfl S. (2020). NHC-gold compounds mediate immune suppression through induction of AHR-TGFβ1 signalling in vitro and in scurfy mice. Communications Biology, 3, 10.
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7

Analyzing NF-κB and Smad Signaling in Ac-HSC

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We further examined the in vitro effects of INT747 and losartan on nuclear factor kappa B (NF‐κB) and mothers against decapentaplegic (Smad) signaling pathways in Ac‐HSC, using western blot analysis. A 50‐μg protein was resolved on 10% acrylamide sodium dodecyl sulfate–polyacrylamide gel electrophoresis gels and transferred to polyvinylidene fluoride membranes by semidry transfer. Membranes were blocked using 5% skim milk in Tris‐buffered saline with Tween 20 (TBST) for 1 hour at room temperature, followed by overnight incubation with 1/1,000 dilution of specific antibodies against phosphorylated (p) ‐NF‐κB (3031), NF‐κB (8242) (Bioss Inc., Woburn, MA), p‐Smad2 (3101), p‐Smad3 (9520), Smad2/3 (3102), and Smad3 (9523) (Cell Signaling Technology Inc., Beverly, MA) at 4 °C; the membranes were then washed in TBST before incubating with horseradish peroxidase‐conjugated secondary antibodies (diluted in 3% [weight per volume] bovine serum albumin in TBST; 1:1,000) for 1 hour at room temperature. After additional washing in TBST, signals were detected using enhanced chemiluminescence, and densitometric scans were quantified using Image J software (NIH).
Namisaki T., Moriya K., Kitade M., Takeda K., Kaji K., Okura Y., Shimozato N., Sato S., Nishimura N., Seki K., Kawaratani H., Takaya H., Sawada Y., Akahane T., Saikawa S., Nakanishi K., Kubo T., Furukawa M., Noguchi R., Asada K., Kitagawa K., Ozutsumi T., Tsuji Y., Kaya D., Fujinaga Y, & Yoshiji H. (2017). Effect of combined farnesoid X receptor agonist and angiotensin II type 1 receptor blocker on hepatic fibrosis. Hepatology Communications, 1(9), 928-945.
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8

Western Blot Analysis of Signaling Pathways

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Western blotting was done as described previously [24 (link)]. The TRPC3 (sc-514670, 1:1000) were purchased from Santa Cruz Biotechnology (Delaware, CA, USA); IL-6 (12,912, 1:1000), AKT (9272,1:1000), ERK (4695,1:1000), p-ERK1/2 (4370, 1:1000), p-SMAD3(9520, 1:1000), NF-KB(8242, 1:1000), p-NF-KB (3033, 1:1000) anti-rabbit IgG-HRP (14,708,1:2000), and anti-mouse IgG-HRP (14,709,1:2000) antibodies were obtained from Cell Signaling Technology (Danvers, MA, USA); IL-8 (500-M08, 1:1000), SMAD3 (25,494-1-AP, 1:1000) and p-Akt (ser473, 66,444-1-IG,1:2000) antibodies were purchased from Proteintech (Chicago, IL, USA).
Xue C., Gao Y., Li X., Zhang M., Yang Y., Han Q., Sun Z., Bai C, & Zhao R.C. (2022). Mesenchymal stem cells derived from adipose accelerate the progression of colon cancer by inducing a MT-CAFs phenotype via TRPC3/NF-KB axis. Stem Cell Research & Therapy, 13, 335.
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