Immunoprecipitation, immunoblotting experiments and Mass Spectrometry analysis were performed as previously described (Lignitto et al., 2019 (link)). Briefly, whole-cell lysates were generated by lysing cells in 50 mM Tris pH 7.5, 150 mM NaCl, 0.5% NP40, 1 mM EDTA, 10% glycerol, protease inhibitor mix (Roche) and phosphatase inhibitor cocktail (Sigma). Total protein amount was measured using a Spectramax spectrophotometer (Molecular Devices, Sunnyvale, CA, USA). Doxycycline (Sigma-Aldrich) was used at 0.1 μg/mL, Cycloheximide (Sigma-Aldrich) was used at 50 μg/mL, MLN4924 (Active Biochem) was used at 2 μM and MG132 (Peptides International) was used at 10 μM. (E/Z)-BCI hydrochloride, PhosSTOP phosphatase inhibitor cocktail tablets were purchased from Sigma-Aldrich. Matrigel Matrix Phenol Red-Free was purchased from Corning. Recombinant Human EGF Protein was obtained from R&D Systems. Purified E1, E2s and ubiquitin were purchased from Boston Biochem (now R&D Systems).
E z bci hydrochloride
Manufactured by Merck Group
The (E/Z)-BCI hydrochloride is a chemical compound used in laboratory settings. It is a salt that can be used as a starting material or intermediate in various chemical reactions and synthesis processes. The compound's core function is to serve as a building block or reactant in experimental and analytical procedures conducted in research and development laboratories.
Automatically generated - may contain errors
Lab products found in correlation
Ubiquitin, by R&D Systems (2 mentions)
Mg132, by Peptide Institute (2 mentions)
Recombinant human egf protein, by R&D Systems (2 mentions)
Matrigel matrix phenol red free, by Corning (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Doxycycline, by Merck Group (2 mentions)
Protease inhibitor mix, by Roche (2 mentions)
Phosphatase inhibitor cocktail, by Merck Group (2 mentions)
Mln4924, by Active Motif (2 mentions)
Spectramax spectrophotometer, by Molecular Devices (2 mentions)
Phosstop phosphatase inhibitor cocktail tablets, by Merck Group (2 mentions)
2 protocols using e z bci hydrochloride
1
Protein Interaction Profiling by Mass Spectrometry
2
Protein Interaction Profiling by Mass Spectrometry
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Immunoprecipitation, immunoblotting experiments and Mass Spectrometry analysis were performed as previously described (Lignitto et al., 2019 (link)). Briefly, whole-cell lysates were generated by lysing cells in 50 mM Tris pH 7.5, 150 mM NaCl, 0.5% NP40, 1 mM EDTA, 10% glycerol, protease inhibitor mix (Roche) and phosphatase inhibitor cocktail (Sigma). Total protein amount was measured using a Spectramax spectrophotometer (Molecular Devices, Sunnyvale, CA, USA). Doxycycline (Sigma-Aldrich) was used at 0.1 μg/mL, Cycloheximide (Sigma-Aldrich) was used at 50 μg/mL, MLN4924 (Active Biochem) was used at 2 μM and MG132 (Peptides International) was used at 10 μM. (E/Z)-BCI hydrochloride, PhosSTOP phosphatase inhibitor cocktail tablets were purchased from Sigma-Aldrich. Matrigel Matrix Phenol Red-Free was purchased from Corning. Recombinant Human EGF Protein was obtained from R&D Systems. Purified E1, E2s and ubiquitin were purchased from Boston Biochem (now R&D Systems).
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