Anti p smad3 c25a9

Manufactured by Cell Signaling Technology

Sourced in China

Anti-p-Smad3 (C25A9) is a primary antibody that detects phosphorylated Smad3 protein. Smad3 is a key intracellular mediator of transforming growth factor-beta (TGF-β) signaling.

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Lab products found in correlation

Ripa lysis buffer, by Beyotime (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Anti bax af0120, by Affinity Biosciences (1 mentions) Anti bcl 2 af6139, by Affinity Biosciences (1 mentions) Anti smad3 c67h9, by Cell Signaling Technology (1 mentions) Chemiscope 6200, by Clinx (1 mentions) Anti gata3 hg3 31, by Santa Cruz Biotechnology (1 mentions) Nupage bis tris gel, by Thermo Fisher Scientific (1 mentions) Immobilon western chemiluminescent hrp substrate, by Merck Group (1 mentions) Anti msc f 20, by Santa Cruz Biotechnology (1 mentions) Anti smad3, by Cell Signaling Technology (1 mentions) Anti β actin sc 47778, by Santa Cruz Biotechnology (1 mentions)

Spelling variants of this product

Smad3 (242 citations) P-Smad3 (240 citations) Anti-Smad3 (93 citations) Anti-p-Smad3 (62 citations)

2 protocols using anti p smad3 c25a9

Protein Expression Analysis in Kidney and Cellular Samples

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Using RIPA lysis buffer (Beyotime), total proteins were extracted from kidneys or cells. The protein concentrations were determined with a BCA protein assay kit (Beyotime). Proteins were transferred onto polyvinylidene difluoride membranes after being separated by 12% SDS-PAGE. Then the membranes were incubated with anti-podocin (BA0290; Boster), anti-p-Smad3 (C25A9; Cell Signaling Technology), anti-Smad3 (C67H9; Cell Signaling Technology), anti-Bax (AF0120; Affinity), anti-Bcl-2 (AF6139; Affinity), and anti-GAPDH (AB0037; Abways, China) antibodies at 4°C overnight. The membranes were treated with the relevant secondary antibody at room temperature for 1 h after being rinsed with Tris-buffered saline with Tween (TBST). The protein bands were depicted with an enhanced ECL kit (Boster) and a chemiluminescence imaging system (ChemiScope 6200; Clinx, China). ImageJ software (NIH, Bethesda, MD, United States) was used to calculate the band gray intensity.

Hu Q.D., Tan R.Z., Zou Y.X., Li J.C., Fan J.M., Kantawong F, & Wang L. (2023). Synergism of calycosin and bone marrow-derived mesenchymal stem cells to combat podocyte apoptosis to alleviate adriamycin-induced focal segmental glomerulosclerosis. World Journal of Stem Cells, 15(6), 617-631.

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Western Blot Analysis of Cellular Signaling Proteins

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0.2–1 × 10⁶ cells were lysed in whole cell extract buffer (50 mM Tris-HCl, pH 7.5, 150 mM NaCl, 0.5% sodium deoxycholate, 0.2 mM EDTA, 10 mM Na₃VO₄, 10% glycerol, protease inhibitors). Proteins were separated by gel electrophoresis using 4–12% NuPAGE Bis-Tris gels (Invitrogen) followed by transfer to nitrocellulose membrane. Membranes were incubated with 5% milk in TBST (0.5 M NaCl, Tris-HCl, pH 7.5, 0.1% (v/v) Tween-20) for 60 min and washed once with TBST. Proteins of interest were detected by incubating membranes over night at 4°C in 5% BSA/TBST with anti-MSC (F-20, Santa Cruz), anti-GATA3 (HG3–31, Santa Cruz) anti-p-Smad3 (C25A9, Cell Signaling), anti-Smad3 (9513, Cell Signaling) or anti-β-Actin (sc-47778, Santa Cruz), washing with TBST three times for 10 min and incubating with horseradish peroxidase–conjugated anti-rabbit or anti-mouse antibody (7074 and 7076, Cell Signaling). Bound antibody was detected by using Immobilon Western chemiluminescent HRP substrate (Millipore).

Wu C., Chen Z., Dardalhon V., Xiao S., Thalhamer T., Liao M., Madi A., Franca R.F., Han T., Oukka M, & Kuchroo V. (2017). The transcription factor musculin promotes the unidirectional development of peripheral Treg cells by suppressing the TH2 transcriptional program. Nature immunology, 18(3), 344-353.

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