Western blot was performed according to previous studies [24 (link), 32 (link)] to assess inflammation (COX-2) and collagen deposition (α-SMA, MMP-9) to further delineate the mechanism of emodin (SMAD3). Mammalian protein lysis buffer (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract the total tissue protein. The same amounts of protein were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and then transferred to polyvinylidene fluoride membranes (EMD Millipore, Billerica, MA, USA). The membranes were incubated in the diluted primary antibodies overnight and stored at 4°C. The primary antibodies were anti-COX-2 antibody (Santa Cruz, 1 : 200 dilution), anti-SMAD3 antibody (Santa Cruz, 1 : 500 dilution), anti-α-SMA antibody (Santa Cruz, 1 : 200 dilution), and anti-beta-actin (β-actin) antibody (Santa Cruz, 1 : 1000 dilution). The membranes were incubated with the secondary antibody followed by horseradish peroxidase (HRP; Santa Cruz). Then, an enhanced chemiluminescence system (EMD Millipore) was used to detect the bands. The intensity of the bands was calculated using Image-Pro Plus 5.0 software (Media Cybernetics Inc., Rockville, MD, USA).
Anti beta actin β actin antibody
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Anti-beta-actin (β-actin) antibody is a primary antibody used to detect the beta-actin protein, which is a ubiquitous and highly conserved cytoskeletal protein found in all eukaryotic cells. The antibody can be used for various applications, including Western blotting, immunohistochemistry, and immunocytochemistry, to study the expression and localization of beta-actin in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Mammalian protein lysis buffer, by Thermo Fisher Scientific (2 mentions)
Enhanced chemiluminescence system, by Merck Group (2 mentions)
Anti cox 2 antibody, by Santa Cruz Biotechnology (2 mentions)
Horseradish peroxidase hrp, by Santa Cruz Biotechnology (2 mentions)
Polyvinylidene fluoride membrane, by Merck Group (2 mentions)
Anti α sma antibody, by Santa Cruz Biotechnology (1 mentions)
Anti smad3 antibody, by Santa Cruz Biotechnology (1 mentions)
Image pro plus 5, by Media Cybernetics (1 mentions)
Anti cox 1 antibody, by Santa Cruz Biotechnology (1 mentions)
Image pro plus software, by Media Cybernetics (1 mentions)
2 protocols using anti beta actin β actin antibody
1
Emodin Mechanism via Inflammation and Fibrosis
2
Western Blot Analyses of Protein Markers
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Western blot analyses were performed as described previously.25 (link) Total protein was extracted by mammalian protein lysis buffer (Thermo Fisher Scientific). Equivalent amounts of protein were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membranes (EMD Millipore, Billerica, MA, USA). The membranes were incubated with the following primary antibodies overnight at 4°C: anti-COX-2 antibody (Santa Cruz Biotechnology Inc, Dallas, TX, USA; 1:200 dilution), anti-COX-1 antibody (Santa Cruz Biotechnology Inc; 1:500 dilution), anti-hypoxia inducible factor-1 alpha (HIF-1α) antibody (Cell Signaling Technology, Inc, Danvers, MA, USA; 1:800 dilution), anti-vascular endothelial growth factor (VEGF) antibody (Santa Cruz Biotechnology Inc; 1:400 dilution), and anti-beta-actin (β-actin) antibody (Santa Cruz Biotechnology Inc; 1:1,000 dilution). The blots were detected by a secondary antibody coupled to horseradish peroxidase (HRP; Santa Cruz Biotechnology Inc) and an enhanced chemiluminescence system (EMD Millipore). The density of specific protein bands was determined by Image-Pro Plus software (v 5.0; Media Cybernetics, Inc, Rockville, MD, USA).
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