Proteins were extracted from cells, liver tissues, and mitochondria using RIPA buffer (Beyotime) with protease and phosphatase inhibitors (Beyotime). Separated by 8−12% SDS‐PAGE, an equal amount of protein was transferred to PVDF membranes (Millipore, Inc.). After blocking with 5% nonfat dry milk in TBST, primary antibodies were added and left to incubate overnight. These antibodies include anti‐PISD (sc‐390070, 1:500; Santa Cruz, CA), anti‐p‐STAT3 (9145, 1:2000; Cell Signaling), anti‐STAT3 (9139, 1:1000; Cell Signaling), anti‐COX IV (4850, 1:1000; Cell Signaling), anti‐Bcl‐2 (AF6139, 1:1000; Affinity), anti‐Bax (AF0120, 1:1000; Affinity), anti‐cleaved caspase‐3 (AF7022, 1:1000; Affinity), anti‐actin (AF7018, 1:6000; Affinity), and OXPHOS rodent WB antibody cocktail (45‐8099, 1:1000; Thermo Fisher Scientific). The blot was then further incubated with HRP‐conjugated secondary antibodies, either goat anti‐rabbit (S0001; Affinity) or goat anti‐mouse (S0002; Affinity). Finally, proteins were visualized with an enhanced chemiluminescence kit (Bio‐Rad).
Anti actin
Manufactured by Affinity Biosciences
Anti-Actin is a laboratory reagent used for the detection and quantification of the actin protein in biological samples. Actin is a fundamental structural protein found in all eukaryotic cells and is involved in various cellular processes. Anti-Actin is a specific antibody that binds to actin, allowing for its identification and measurement in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Complete protease inhibitor cocktail, by Roche (2 mentions)
Anti cox 4, by Cell Signaling Technology (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa buffer, by Beyotime (1 mentions)
Protease and phosphatase inhibitor, by Beyotime (1 mentions)
Anti p stat3, by Cell Signaling Technology (1 mentions)
Enhanced chemiluminescence kit, by Bio-Rad (1 mentions)
Anti bax, by Affinity Biosciences (1 mentions)
Anti cleaved caspase 3, by Affinity Biosciences (1 mentions)
Anti stat3, by Cell Signaling Technology (1 mentions)
Oxphos rodent wb antibody cocktail, by Thermo Fisher Scientific (1 mentions)
Anti bcl 2 af6139, by Affinity Biosciences (1 mentions)
Protein assay, by Bio-Rad (1 mentions)
Anti ha, by Santa Cruz Biotechnology (1 mentions)
Anti gfp, by Merck Group (1 mentions)
3 protocols using anti actin
1
Protein Expression Analysis in Tissue Samples
2
JAZ9 Protein Degradation Assay
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Six-day-old seedlings grown on 1/2 MS medium under LD conditions were placed in darkness for 12 h before treatment. Then, seedlings were treated with darkness, red light and white light for 24 h separately. Samples were harvested at indicated time, total proteins were extracted using extraction buffer containing 100 mM Tris–HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 1% SDS, 1 × complete protease inhibitor cocktail (Roche) and 50 μM MG132. Protein concentration was determined by the Bio-Rad protein assay. The anti-HA (Santa Cruz) and anti-Actin (Affinity) were used to detect HA-JAZ9 and Actin, respectively. The immunoblot bands were quantified by ImageJ.
3
Extracting Proteins from 35S:SE-GFP Seedlings
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Fourteen-day-old 35SPro:SE-GFP transgenic seedings were harvested after treatment with NaCl and total proteins were extracted using the protein extraction buffer containing 100 mM Tris–HCl, pH 7.5, 100 mM NaCl, 5 mM EDTA, 1% SDS, 1×complete protease inhibitor cocktail (Roche) and 50 μM MG132. The anti-GFP (Sigma–Aldrich) and anti-Actin (Affinity) antibody were used to detect SE-GFP and Actin, respectively.
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