Anti smad2 3 3102

Manufactured by Cell Signaling Technology

Sourced in United States

Anti-SMAD2/3 (3102) is a laboratory reagent used for the detection and measurement of SMAD2 and SMAD3 proteins in cellular samples. It is a primary antibody that specifically binds to these proteins, allowing for their identification and quantification through various analytical techniques.

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Lab products found in correlation

Image studio lite software, by LI COR (1 mentions) Anti tubulin, by Merck Group (1 mentions) Draq5 fluorescent probe, by Thermo Fisher Scientific (1 mentions) Anti β tubulin, by Merck Group (1 mentions) Anti cd68 ebm11, by Agilent Technologies (1 mentions) Cd140a pe, by BD (1 mentions) Cell culture media, by Lonza (1 mentions) Il 1β, by Thermo Fisher Scientific (1 mentions) Anti α sma, by Merck Group (1 mentions) Trypsin, by Lonza (1 mentions) Phosphate buffered saline (pbs), by Lonza (1 mentions) Cd56 apc, by BD (1 mentions) Ab31013, by Abcam (1 mentions) Ab186838, by Abcam (1 mentions) Ab92486, by Abcam (1 mentions) Anti ctgf sc 14939, by Santa Cruz Biotechnology (1 mentions) Sc 8782, by Santa Cruz Biotechnology (1 mentions) Anti gapdh antibody sc 47724, by Santa Cruz Biotechnology (1 mentions) Gel pro analysis software, by Media Cybernetics (1 mentions) Anti cd31 antibody 550 274, by BD (1 mentions)

4 protocols using anti smad2 3 3102

Protein Expression Analysis in Ovine Cells

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Cells were lysed, fractionated by SDS-PAGE and transferred to 0.45 PVDF membranes as described [13 ]. Blots were probed with goat anti-human CD31 (1:300) (M-20, Santa Cruz Biotechnology), goat anti-human VE-cadherin (1:300) (C-19, Santa Cruz Biotechnology, murine anti-human α-SMA (1:2000) (Sigma, mAb clone 1A4) and anti-tubulin (Sigma). Two different anti-p-ERK antibodies were used. The first was a murine mAb anti-phospho-44/42 MAPK (Erk1/2) (Thr202/Tyr204) and the second was a rabbit polyclonal anti-phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204). Total ERK was detected using anti-p44/42 MAPK (ERK) mAb. Anti-phosphorylated-SMAD3 (p-SMAD3) (Ser423/425(clone C25A9) and anti-SMAD2/3 (3102), and all ERK pathway antibodies, were from Cell Signaling Technology. All antibodies were shown to cross-react with their ovine homologs. Band intensities on the western blots were quantified using Image Studio Lite software (LI-COR).

Wylie-Sears J., Levine R, & Bischoff J. (2014). Losartan Inhibits Endothelial-to-Mesenchymal Transformation in Mitral Valve Endothelial Cells by Blocking Transforming Growth Factor-β-induced Phosphorylation of ERK. Biochemical and biophysical research communications, 446(4), 870-875.

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Immunofluorescence and Western Blot Analysis

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Cell culture media, serum, phosphate-buffered saline and trypsin were purchased from Lonza (Verviers, Belgium) and cell culture reagents were from Sigma-Aldrich Chimie (Saint-Quentin Fallavier, France).
Human antibodies for immunofluorescence analysis were purchased as indicated: anti-αSMA (#A5228) from Sigma-Aldrich, anti-CD68 (#EBM11) from Dako (Glostrup, Denmark), anti-MR (#ab8918) from Abcam (Cambridge, United-Kingdom), anti-CD140a (#3174 T) from Cell Signaling (Ozyme, St Quentin en Yvelines, France). Human antibodies for western blot analysis were purchased as indicated: anti-phospho-Smad2 (Ser465/467, #3101) and anti-Smad2/3 (#3102) from Cell Signaling, and anti-β-tubulin from Sigma-Aldrich. Human antibodies against CD56-APC (#341027) and CD140a-PE (#556002) were purchased from BD-Biosciences (Le Pont de Claix, France). Human recombinant IL-4 (#200-04) and IL-1β (#200-01B) were purchased from Peprotech (Neuilly-Sur-Seine, France) and used at 15 ng/ml. SB431542 (#S4317) was purchased from Sigma-Aldrich Chimie and used at 5 µM. DRAQ5 fluorescent probe (#62254) was purchased from Thermofisher Scientific and used at 20 µM.

Moratal C., Raffort J., Arrighi N., Rekima S., Schaub S., Dechesne C.A., Chinetti G, & Dani C. (2018). IL-1β- and IL-4-polarized macrophages have opposite effects on adipogenesis of intramuscular fibro-adipogenic progenitors in humans. Scientific Reports, 8, 17005.

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Protein Expression Analysis of TGF-β Signaling

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Total protein was extracted from treated cells using RIPA buffer. Equal amounts of cell extracts were fractionated by SDS-PAGE and analyzed by Western blotting and visualized by enhanced chemiluminescence. The following primary antibodies were employed: antipro-COL1A1 (SC-8782, Santa Cruz, Dallas, TX, USA), anti-TGF-β1 (ab92486, Abcam, Cambridge, MA, USA), anti-TGF-β receptor I antibody (ab31013, Abcam, Cambridge, MA, USA), anti-TGF-β receptor II (ab186838, Abcam, Cambridge, MA USA), antiphospho-Smad2/Smad3 (8828, Cell Signaling, Danvers, MA, USA), anti-Smad2/3 (3102, Cell Signaling, Danvers, MA, USA), anti-CTGF (SC-14939, Santa Cruz, California) and anti-GAPDH antibody (sc-47724, Santa Cruz, Dallas, TX, USA) The quantification of protein bands was carried out using GelPro Analysis Software (Media Cybernetics).

Wang P., Han J., Wei M., Xu Y., Zhang G., Zhang H., Shi L., Liu X., Hamblin M.R, & Wang X. (2018). Remodeling of dermal collagen in photoaged skin using low-dose 5-aminolevulinic acid photodynamic therapy occurs via the transforming growth factor-β pathway. Journal of biophotonics, 11(6), e201700357.

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Differentiation of Mouse Embryonic Stem Cells

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J1 mESC cell line was purchased from American Type Culture Collection. The anti-VE-cadherin antibody (sc-9989), anti-NANOG (sc-134218), anti-TGFβRI (sc-398, sc-33933), anti-TGFβRII (sc-400, sc-17792), anti-PAR1 (sc-5605), anti-6×His rabbit antibody (sc-803), anti-6×His mouse antibody (sc-8036), and anti-α-actin (sc-32251) antibodies were from Santa Cruz Biotechnology. Anti-VE-cadherin antibody (AF1002), recombinant mouse TGF-β1 (7666-MB-005), and Mouse TER-119 antibody (MAB1125, targets erythrocytes) were from R&D Systems. Alexa Fluor 633 goat anti-rat immunoglobulin G (IgG) (H+L) (A21092, Life Technologies) was used as secondary antibody for detecting TER-119. Collagen IV-coated 6-well plate (354428) and Matrigel were from BD Biosciences. Anti-CD31 antibody (550274) was purchased from BD Biosciences/Pharmingen. Anti-FLK1 antibody (136404) was purchased from Biolegend. Anti-SMAD2/3 (3102), anti-phospho-SMAD2 (3108), and anti-FLK1 (2479) antibodies were from Cell Signaling Technology. The PAR1 agonist peptide (TFLLRNPNDK-NH₂) was synthesized and purified at the Research Resource Center at the University of Illinois, Chicago.

Gong H., An S., Sassmann A., Liu M., Mastej V., Mittal M., Zhang W., Hong Z., Offermanns S., Rehman J, & Malik A.B. (2016). PAR1 Scaffolds TGFβRII to Downregulate TGF-β Signaling and Activate ESC Differentiation to Endothelial Cells. Stem Cell Reports, 7(6), 1050-1058.

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