Total protein was extracted from cells by RIPA lysis (Beyotime Institute of Biotechnology, Jiangsu, China) containing 1% protease/phosphatase inhibitor cocktail (ComWin Biotech Corporation, Beijing, China) and then measured by Pierce BCA Protein Assay Kit (Thermo Fisher Scientific, USA) to determine the protein concentration. Protein lysates were loaded and separated in 4–20% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) gel (Sigma, USA) and transferred to polyvinylidene difluoride (PVDF) membranes after electrophoresis (EMD Millipore, Billerica, MA, USA). After blocking with 5% nonfat dry milk for 1 h at room temperature, the membranes were washed and incubated with Notch1 and glyceraldehyde 3-phosphate dehydrogenase (GADPH) primary antibody (20,687-1-AP and 10,494-1-AP, Proteintech Group Incorporation, IL, USA) in dilutions of 1:1000 and 1:3000 at 4°C overnight. The next day, the membranes were washed and incubated with horseradish peroxidase-labeled secondary antibody (SA00001-2, Proteintech Group Incorporation, IL, USA) at a 1:5000 dilution. Then, the antibody-labeled bands were detected by an enhanced chemiluminescence detection system (ECL, BestBio Pharmacia Biotech). The results were quantified with Quantity One Analysis Software (Bio-Rad Laboratories, Hercules, CA, 210 USA).
Protease phosphatase inhibitor cocktail
Manufactured by CoWin Biotech
Sourced in China
The Protease/phosphatase inhibitor cocktail is a laboratory reagent designed to inhibit the activities of proteases and phosphatases. It is commonly used in protein extraction and purification processes to prevent the degradation of target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Tumor necrosis factor alpha tnf α, by Thermo Fisher Scientific (2 mentions)
Phospho nf κb p65 ser536, by Cell Signaling Technology (2 mentions)
Phospho iκbα ser32, by Cell Signaling Technology (2 mentions)
Uronic acid, by Yuanye Bio-Technology (2 mentions)
Bicinchoninic acid bca protein assay, by CoWin Biotech (2 mentions)
Anti β actin antibodies, by CoWin Biotech (2 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by CoWin Biotech (2 mentions)
Lipopolysaccharides (lps), by Solarbio (2 mentions)
Streptozotocin (stz), by Merck Group (2 mentions)
Nf κb p65, by Cell Signaling Technology (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ripa lysis, by Beyotime (1 mentions)
Quantity one analysis software, by Bio-Rad (1 mentions)
Polyvinylidene difluoride pvdf membrane, by Merck Group (1 mentions)
3 protocols using protease phosphatase inhibitor cocktail
1
Western Blot Analysis of Notch1 Protein
2
Antidiabetic and Anti-inflammatory Effects
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The reference standard of glucose and uronic acid were provided by Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Moreover, metformin hydrochloride tablets were purchased from Sino-American Shanghai Squibb Pharmaceuticals Ltd. (Shanghai, China). STZ was obtained from Sigma-Aldrich (Sacramento, CA, USA). All test assay kits and enzyme-linked immunosorbent assay (ELISA) kits were supplied by NanJing JianCheng Bioengineering Institute (Nanjing, China). Tumor necrosis factor-alpha (TNF-α) was obtained from PeproTech Inc. (Cranbury, NJ, USA). Lipopolysaccharides (LPS) were provided by Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). NF-κB p65 (1:2,000), phospho-NF-κB p65(Ser536; 1:2,000), and phospho-IκBα (Ser32; 1:2,000) were purchased from Cell Signaling Technology (Beverly, MA, USA). The radioimmunoprecipitation assay (RIPA) lysis buffer with protease/phosphatase inhibitor cocktail, bicinchoninic acid (BCA) protein assay, anti-β-actin antibodies (1:2,000), and horseradish peroxidase (HRP)-conjugated secondary antibodies was purchased from Beijing ComWin Biotech Co., Ltd. (Beijing, China). All other chemical reagents were of analytical grade.
3
Metformin Attenuates Inflammatory Response
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The reference standard of glucose and uronic acid were provided by Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Moreover, metformin hydrochloride tablets were purchased from Sino-American Shanghai Squibb Pharmaceuticals Ltd. (Shanghai, China). STZ was obtained from Sigma-Aldrich (Sacramento, CA, USA). All test assay kits and enzyme-linked immunosorbent assay (ELISA) kits were supplied by NanJing JianCheng Bioengineering Institute (Nanjing, China). Tumor necrosis factor-alpha (TNF-α) was obtained from PeproTech Inc. (Cranbury, NJ, USA). Lipopolysaccharides (LPS) were provided by Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). NF-κB p65 (1:2,000), phospho-NF-κB p65(Ser536; 1:2,000), and phospho-IκBα (Ser32; 1:2,000) were purchased from Cell Signaling Technology (Beverly, MA, USA). The radioimmunoprecipitation assay (RIPA) lysis buffer with protease/phosphatase inhibitor cocktail, bicinchoninic acid (BCA) protein assay, anti-β-actin antibodies (1:2,000), and horseradish peroxidase (HRP)-conjugated secondary antibodies was purchased from Beijing ComWin Biotech Co., Ltd. (Beijing, China). All other chemical reagents were of analytical grade.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!