Cell lysates were prepared by briefly sonicating cells in modified RIPA buffer (10 mM Na2HPO4, 1.8 mM KH2PO4, 137 mM NaCl, 2.7 mM KCl, 0.5% deoxycholate, 1% Nonidet P-40, 0.3% SDS) plus cocktails of protease inhibitors and phosphatase inhibitors (Sigma, St. Louis, MO, USA). After protein concentrations were determined by BCA protein assay kit (Thermo Fisher Scientific), cell lysates were subjected to conventional SDS-PAGE and immunoblot analysis procedures. Finally, protein bands were detected by enhanced chemiluminescence (Luminata™ Crescendo Western HRP Substrate, EMD Millipore).
Cocktails of protease inhibitors and phosphatase inhibitors
Manufactured by Merck Group
Sourced in United States
Cocktails of protease inhibitors and phosphatase inhibitors are a type of lab equipment used in various research applications. These cocktails contain a combination of chemical compounds that inhibit the activity of proteases and phosphatases, which are enzymes involved in various biological processes. The core function of these cocktails is to provide a tool for researchers to study and manipulate the activities of these enzymes in their experimental systems.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using cocktails of protease inhibitors and phosphatase inhibitors
1
Cell Lysis and Immunoblot Analysis
2
Cell Lysis and Immunoblot Analysis
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Cell lysates were prepared by briefly sonicating cells in lysis buffer [18 (link)] plus cocktails of protease inhibitors and phosphatase inhibitors (Sigma-Aldrich, St. Louis, MO). The protein concentrations of cell lysates were determined by BCA protein assay kit (Thermo Fisher Scientific), and immunoblot analyses were performed according to the procedure described previously [17 (link)]. The protein bands were detected by enhanced chemiluminescence (LuminataTM Crescendo Western HRP Substrate, EMD Millipore). The antibodies used were listed in Additional file 1 : Table S1.
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