The concentration of the extracted protein was measured using a BCA protein assay kit (Beyotime, China) according to the manufacturer's protocol. After SDS-polyacrylamide gel electrophoresis, the separated protein was transferred onto polyvinylidene difluoride membranes (Millipore, USA) at 80 V for 1.5 h. The protein was then subjected to immune blot analysis with specific antibodies for ITGBL1 (Abclonal, China), Smad2 (Abclonal), p-Smad2 (Abclonal), Smad3 (Abclonal), p-Smad3 (Abclonal), Smad7 (Abclonal), JDP2 (Affinity, China), TGF-β1 (Abclonal), Ki67 (Abclonal), MMP2 (Proteintech, China), and β-actin (Santa Cruz, USA). All antibodies were diluted to 1:1000, except JDP2, which was diluted to 1:2000. In addition, the secondary antibody reagent in this study was IgG-HRP (Beyotime, China) with 1:5000 dilutions.
Smad7
Manufactured by ABclonal
Sourced in China
Smad7 is a lab equipment product used for research purposes. It functions as a regulatory protein involved in the transforming growth factor-beta (TGF-beta) signaling pathway.
Automatically generated - may contain errors
Lab products found in correlation
Smad3, by ABclonal (2 mentions)
Polyvinylidene difluoride membrane, by Merck Group (1 mentions)
Tgf β1, by ABclonal (1 mentions)
Smad2, by ABclonal (1 mentions)
P smad2, by ABclonal (1 mentions)
P smad3, by ABclonal (1 mentions)
Igg hrp, by Beyotime (1 mentions)
β actin, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (1 mentions)
Sds page, by Bio-Rad (1 mentions)
Col3a1, by ABclonal (1 mentions)
Phospho smad3, by ABclonal (1 mentions)
α sma, by ABclonal (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Keygen Biotech (1 mentions)
Gapdh, by ABclonal (1 mentions)
Col1a1, by ABclonal (1 mentions)
2 protocols using smad7
1
Protein Expression Analysis by Western Blot
2
Protein Expression Analysis in Cardiac Tissues
Check if the same lab product or an alternative is used in the 5 most similar protocols
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NRCFs or left ventricular tissues were homogenized in lysis buffer containing PMSF (Keygen). To obtain proteins of high purity, lysates were centrifuged at 12,000
g for 15 min. The bicinchoninic acid protein assay kit (Thermo Fisher) was used to measure the protein concentration. Each sample containing 20 μg of protein was separated by 10% SDS-PAGE (Bio-Rad) and then transferred onto a PVDF membrane (Millipore, Billerica, USA). Thereafter, the proteins were probed with primary antibodies against α-SMA (1:1000; Abclonal, Wuhan, China), CTGF (1:500; Abclonal), Col1a1 (1:1000; Abclonal), Col3a1 (1:1000; Abclonal), Smad7 (1:500; Abclonal), Smad3 (1:500; Abclonal), phospho-Smad3 (1:500; Abclonal), and GAPDH (1:2000; Abclonal). Subsequently, the membranes were incubated with a secondary antibody, namely, HRP-linked goat anti-rabbit IgG (1:5000; Abclonal). Finally, the signals were detected using an ECL kit (Tanon, Shanghai, China) on an imaging system (Tanon), and the blots were quantified with ImageJ software.
g for 15 min. The bicinchoninic acid protein assay kit (Thermo Fisher) was used to measure the protein concentration. Each sample containing 20 μg of protein was separated by 10% SDS-PAGE (Bio-Rad) and then transferred onto a PVDF membrane (Millipore, Billerica, USA). Thereafter, the proteins were probed with primary antibodies against α-SMA (1:1000; Abclonal, Wuhan, China), CTGF (1:500; Abclonal), Col1a1 (1:1000; Abclonal), Col3a1 (1:1000; Abclonal), Smad7 (1:500; Abclonal), Smad3 (1:500; Abclonal), phospho-Smad3 (1:500; Abclonal), and GAPDH (1:2000; Abclonal). Subsequently, the membranes were incubated with a secondary antibody, namely, HRP-linked goat anti-rabbit IgG (1:5000; Abclonal). Finally, the signals were detected using an ECL kit (Tanon, Shanghai, China) on an imaging system (Tanon), and the blots were quantified with ImageJ software.
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