The prepared protein samples were quantified using the bicinchoninic acid (BCA) protein assay kit (CWBIO Corporation) and protein extraction kit (Beijing ComWin Biotech Co., Ltd.). Next, Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) was used to divide and transfer the protein samples to the Polyvinylidene Fluoride (PVDF) membranes. The cells were incubated in 5% skim milk for 1 hour to block non-specific binding. Subsequently, the membranes were probed with primary antibodies at 4 ℃ overnight and with horseradish peroxidase–linked anti-rabbit (cat. no. ab6721; 1:2000; Abcam) secondary antibody on the following day. The enhanced chemiluminescence detection system was used to visualize the protein bands, and band intensity was determined using LI-COR Image Studio v5.2.5 Software (LI-COR Biosciences). The value was normalized to GAPDH. Anti-TGFβ1 (cat. No. 21898-1-AP; 1:1,000; Proteintech), anti- alpha smooth muscle actin (α-SMA; cat. No. #19245; 1:1,000; Cell Signaling Technology), and anti-GAPDH (cat. No. ab9485; 1:2,500; Abcam) were used.
Anti alpha smooth muscle actin α sma
Manufactured by Proteintech
Anti-alpha smooth muscle actin (α-SMA) is an antibody that recognizes the alpha isoform of smooth muscle actin, a key structural protein found in smooth muscle cells. This antibody can be used to detect and localize smooth muscle actin in a variety of cell and tissue types.
Automatically generated - may contain errors
Lab products found in correlation
Image studio v5, by LI COR (1 mentions)
Anti gapdh, by Cell Signaling Technology (1 mentions)
Protein extraction kit, by CoWin Biotech (1 mentions)
Azithromycin, by Merck Group (1 mentions)
Anti loxl2, by Abcam (1 mentions)
Anti collagen 1, by Proteintech (1 mentions)
Anti α tubulin, by Proteintech (1 mentions)
Anti vimentin, by Proteintech (1 mentions)
Anti p jnk, by Proteintech (1 mentions)
Anti smad2, by Cell Signaling Technology (1 mentions)
Anti smad3, by Cell Signaling Technology (1 mentions)
Anti c jun, by Proteintech (1 mentions)
Anti p smad3, by Cell Signaling Technology (1 mentions)
Anti gapdh, by Proteintech (1 mentions)
Anti jnk, by Proteintech (1 mentions)
Anti tgf β1, by Proteintech (1 mentions)
2 protocols using anti alpha smooth muscle actin α sma
1
Quantification and Analysis of Protein Expression
2
Fibrosis Signaling Pathway Modulation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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BLM and the JNK inhibitor (SP600125) were obtained from Selleck China Inc. (Shanghai, China). TGF-β1 was purchased from PeproTech China Inc. (Suzhou, China). Azithromycin was obtained from Sigma-Aldrich Inc. (Shanghai, China). The primary antibodies we used are as follows: anti-vimentin (Proteintech, 60330-1-Ig), anti-alpha-smooth muscle actin (α-SMA) (Proteintech, 14395-1-AP), anti-Collagen 1 (Proteintech, 14695-1-AP), anti-LOX (Proteintech, 17958-1-AP), anti-LOXL2 (Abcam, 96233), anti-TGF-β1 (Proteintech, 21898-1-AP), anti-Smad2 (Cell Signaling Technology, 5339), anti-Smad3 (Cell Signaling Technology, 9523), anti-phospho (P)-smad2 (Cell Signaling Technology, 3108), anti-P-smad3 (Cell Signaling Technology, 9520), anti-JNK (Proteintech, 66210-1-Ig), anti-c-Jun (Proteintech, 66313-1-Ig), anti-P-JNK (Proteintech, 80024-1-RR), anti-P-cJun (Proteintech, 28891-1-AP), anti-α-tubulin (Proteintech, 66031-1-Ig), and anti-GAPDH (Proteintech, 60004-1-Ig). The dilution ratio of all antibodies was 1:1000.
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