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Smad2 3103

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Smad2 (#3103) is a primary antibody that recognizes endogenous levels of total Smad2 protein. Smad2 is a central mediator of transforming growth factor-beta (TGF-beta) signaling.

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Lab products found in correlation

Erk1 2 9102, by Cell Signaling Technology (2 mentions) Smad3 9523, by Cell Signaling Technology (2 mentions) E cadherin, by BD (2 mentions) P smad2 3108, 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) P smad3 9520, 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) Phosstop phosphatase inhibitor cocktail, by Roche (1 mentions) G8795, by Merck Group (1 mentions) Protease inhibitor cocktail, by Roche (1 mentions) Control mirna, by Bioneer (1 mentions)

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Smad2 (224 citations)

4 protocols using smad2 3103

1

Extraction and Western Blot Analysis of Phospho-Smad Proteins

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Proteins from tail tendons were extracted from frozen tissue with ice-cold cell lysis buffer (20 mM Tris/HCl pH 7.6, 150 mM NaCl, 1 mM ethylene diamine tetra-acetic acid (EDTA), 1% (v/v) Igepal CA-630, 50 mM sodium fluoride) containing complete, mini EDTA-free protease inhibitor cocktail and PhosSTOP phosphatase inhibitor cocktail (Roche). Lysates were cleared by centrifugation at 10 000 × g, at 4oC. Commercially available antibodies used to detect phospho-Smad1/5 (#9516), Smad1 (#6933), Smad5 (#9517), phospho-Smad2 (#3108) and Smad2 (#3103) were purchased from Cell Signaling Technologies. GAPDH was detected using a monoclonal antibody from Sigma (G8795).
Yeung C.Y., Gossan N., Lu Y., Hughes A., Hensman J.J., Bayer M.L., Kjær M., Kadler K.E, & Meng Q.J. (2014). Gremlin-2 is a BMP antagonist that is regulated by the circadian clock. Scientific Reports, 4, 5183.
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2

Quantification of Smooth Muscle Markers

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Human recombinant platelet derived growth factor-BB (PDGF-BB) was purchased from Sigma (St. Louis, MO, USA). MiRNA-18a-5p and the miRNA control were purchased from Bioneer Co. (Daejeon, Korea).
Gapdh (sc-32233), syndecan4 (sc-15350), smooth muscle (SM) 22α (sc-373928), and SM α-actin (sc-130617) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Smad2 (#3103) was purchased from Cell Signaling Technology (Danvers, MA, USA).
Kee H.J., Kim G.R., Cho S.N., Kwon J.S., Ahn Y., Kook H, & Jeong M.H. (2014). miR-18a-5p MicroRNA Increases Vascular Smooth Muscle Cell Differentiation by Downregulating Syndecan4. Korean Circulation Journal, 44(4), 255-263.
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3

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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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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