Dulbecco’s modified Eagles’ medium (DMEM), penicillin/streptomycin, trypsin/EDTA, fetal bovine serum (FBS), KGM-SFM medium, and phosphate-buffered saline (PBS) were from Gibco (Life Technologies, USA). Arecoline, melatonin and MTT were from Sigma (Sigma-Aldrich Chemical Company, St. Louis, MO, USA). ELISA kits for MMP-9 were from PeproTech (PeproTech, Inc., Rocky Hill, NJ, USA). Catalase, SB431542, 5Z-7-Oxozeaenol, PD153035, AG490, U0126, LY294002, and aspirin were purchased from Tocris. Phospho-TAK1 (p-TAK1) antibody was from Abcam (ab79583). Antibodies for TGF-β1, and GAPDH were from Santa Cruz. P-Smad2 antibody was from Cell Signaling Technology. PBL extract and ANE were prepared as before [8 (link), 12 (link), 13 (link), 21 (link)]. HC was synthesized as previously [22 (link)].
P smad2 antibody
Manufactured by Cell Signaling Technology
Sourced in United States
The P-Smad2 antibody is a specific antibody that recognizes the phosphorylated form of the Smad2 protein. Smad2 is a key mediator of transforming growth factor-beta (TGF-β) signaling pathway. The phosphorylation of Smad2 is a critical step in the activation of this signaling cascade.
Automatically generated - may contain errors
Lab products found in correlation
Aspirin, by Bio-Techne (1 mentions)
Melatonin, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
5z 7 oxozeaenol, by Bio-Techne (1 mentions)
Arecoline, by Merck Group (1 mentions)
Pd153035, by Bio-Techne (1 mentions)
Ag490, by Bio-Techne (1 mentions)
Ly294002, by Bio-Techne (1 mentions)
U0126, by Bio-Techne (1 mentions)
Sb431542, by Bio-Techne (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Infrared labeled secondary antibody, by LI COR (1 mentions)
Odyssey clx infrared imaging system, by LI COR (1 mentions)
Tsp 1 antibody, by Cell Signaling Technology (1 mentions)
Tgf β1 antibody, by Cell Signaling Technology (1 mentions)
β actin antibody, by Merck Group (1 mentions)
3 protocols using p smad2 antibody
1
Molecular Mechanisms Regulating Cell Signaling
2
Protein Expression Quantification
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The primary antibodies were used as follows: TSP-1 antibody (1:1000, Cell signaling technology, USA), GAP-43 antibody (1:1000, Cell signaling technology, USA), TGF-β1 antibody (1:1000, Cell signaling technology, USA), smad2 antibody (1:1000, Cell signaling technology, USA), p-Smad2 antibody (1:1000, Cell signaling technology, USA), and β-actin antibody (1:5000; Sigma, USA). After several washes with TBS-T, blots were incubated with infrared labeled secondary antibody (Li-COR Biosciences). Immunoblot bands were visualized and analyzed by using Odyssey CLx infrared imaging system (LI-COR® Biosciences, USA).
3
Quantifying Phosphorylated SMAD2 in Palate
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Cells containing phosphorylated-SMAD2 were detected in paraffin sections by using Psmad2 antibody (1:20, Cell Signaling Technology, USA) (Fujita et al., 2012) . Slides were first deparaffinized and then rehydrated (see section 3.3.3) . Antigen retrieval was done in a pressure cocker and a blocking solution was applied (see section 3.3.4) . Then primary antibodies were diluted with blocking solution, Psmad2 (1:20, Cell Signaling Technology, USA) and Ecadherin Nine sections obtained from each sample for each secondary palate location (Anterior, Middle and Posterior) were analyzed, and average values were calculated for ten samples in each group. These mean values and the standard deviations were used for one-way ANOVA that compared Psmad2 positive cells in the wild type, K14-SMAD2/ TGF-β3 (+/-) and K14-SMAD2/TGF-β3 (-/-) mice groups.
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