Nuclear and cytoplasmic protein extracts were prepared as previously described using nuclear and cytoplasmic protein extraction reagent kits (Beyotime Biotechnology) following the manufacturer’s instructions.23 (link) The protein concentration was determined with bicinchoninic acid assay kits (Thermo Fisher Scientific). The proteins in samples of equal concentration were separated by 10% and 15% sodium dodecyl-sulfate polyacrylamide gel electrophoresis gels and transferred to a Nitrocellulose membrane. After blocking, the membranes were incubated with primary antibodies at 4°C overnight. The next day, the membranes was incubated with the anti-rabbit secondary antibody for 1 h at room temperature. The protein bands were detected by chemiluminescence and the intensity was calculated by densitometry. Primary antibodies and working dilutions are detailed in Supplementary Table 1 .
Nuclear and cytoplasmic protein extraction reagent kits
Manufactured by Beyotime
Sourced in China
The nuclear and cytoplasmic protein extraction reagent kits are designed to isolate and purify nuclear and cytoplasmic proteins from cells or tissues. The kits contain the necessary reagents and buffers to effectively separate these protein fractions for further analysis or downstream applications.
Automatically generated - may contain errors
Lab products found in correlation
Bicinchoninic acid assay kit, by Thermo Fisher Scientific (1 mentions)
α bgt, by Merck Group (1 mentions)
Anti β actin monoclonal antibody, by Cell Signaling Technology (1 mentions)
Anti nf κb p65, by Cell Signaling Technology (1 mentions)
Lps escherichia coli 0111 b4, by Merck Group (1 mentions)
Anti tnf α, by Cell Signaling Technology (1 mentions)
Anti il 6, by Cell Signaling Technology (1 mentions)
Anti lamin b, by Cell Signaling Technology (1 mentions)
Anti il 1β, by Cell Signaling Technology (1 mentions)
Dexamethasone (dex), by Jiangsu Hengrui Medicine (1 mentions)
2 protocols using nuclear and cytoplasmic protein extraction reagent kits
1
Nuclear and Cytoplasmic Protein Extraction
2
Inflammatory Cytokine Regulation Assay
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Dex was obtained from Jiangsu Hengrui Medicine Co., Ltd. (Jiangsu, China). Mouse TNF-α, IL-6, and IL-1β assay kits were purchased from Shanghai Lengton Bioscience Co., Ltd. (Shanghai, China). LPS (Escherichia coli 0111: B4) and α-Bgt were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Nuclear and cytoplasmic protein extraction reagent kits were obtained from Beyotime Institute of Biotechnology (Shanghai, China). Anti-TNF-α, anti-IL-6, anti-IL-1β, anti-p-IκB p65, anti-IκB p65, anti-NF-κB p65, anti-Lamin B, and anti-β-actin monoclonal antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). All other chemicals were reagent grade.
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