vSMC or cleaned aortas were incubated on ice with lysis buffer supplemented with proteases and phosphatases inhibitors cocktails (Sigma) and sodium orthovanadate. Lysates were subjected to SDS‐PAGE, transferred to nitrocellulose membranes and incubated with specific antibodies. Immune complexes were detected with appropriate secondary antibodies and enhanced chemiluminescence reagent (ECL plus; GE Healthcare). Protein band intensities were quantified using ImageJ Software (NIH software).
Proteases and phosphatases inhibitors cocktails
Manufactured by Merck Group
Proteases and phosphatases inhibitors cocktails are a range of laboratory products provided by Merck Group. These cocktails contain a mixture of compounds that can effectively inhibit the activity of proteases and phosphatases, which are important enzymes in biological research and analysis. The core function of these cocktails is to help preserve the integrity of protein samples by preventing unwanted degradation during sample preparation and processing.
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Lab products found in correlation
3 protocols using proteases and phosphatases inhibitors cocktails
1
Western Blot Protein Analysis
2
Protein Extraction from Tissue Samples
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Several milligrams of tissue were pulverized and then were homogenized in 10 mmol/L Tris-HCl, pH 7.5; 1 mmol/L MgCl2, 1 mmol/L EGTA, 10% glycerol, 0.5% CHAPS and proteases and phosphatases inhibitors cocktails (Sigma) at 4 °C using an Ultra-turrax dispersing instrument (Ika). The extracts were vortexed for 30 min at 4 °C and after centrifuging for 15 min at 13000 rpm, the supernatants were stored at -80 °C. Protein levels were determined with Bradford reagent (Bio-Rad).
3
Protein Extraction from Cells
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Cells were homogenized in a medium containing 10 mmol/L Tris-HCl, pH 7.5; 1 mmol/L MgCl2, 1 mmol/L EGTA, 10% glycerol, 0.5% CHAPS and proteases and phosphatases inhibitors cocktails (Sigma). The extracts were vortexed for 30 min at 4 °C and after centrifuging for 15 min at 13000 rpm, the supernatants were stored at -20 °C. Protein levels were determined with Bradford reagent (Bio-Rad).
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