Author: Clayton M. Carey; Sarah E. Apple; Zoe A. Hilbert; Michael S. Kay; Nels C. Elde
Title: Conflicts with diarrheal pathogens trigger rapid evolution of an intestinal signaling axis Document date: 2020_3_30
ID: ju826pao_27
Snippet: Western blotting 293T cells expressing GC-C were grown to confluence in 24-well plates following immunoblot analysis. To avoid aggregation, cells were lysed directly in 2x Laemmli buffer containing 8 M urea (Sigma), 3 M thiourea (Sigma), and β-mercaptoethanol for 30 min at room temperature prior to analysis. Total protein was resolved by Mini-PROTEAN GTX polyacrylamide gel electrophoresis (Bio-rad). Proteins were detected using anti-HA (Covance .....
Document: Western blotting 293T cells expressing GC-C were grown to confluence in 24-well plates following immunoblot analysis. To avoid aggregation, cells were lysed directly in 2x Laemmli buffer containing 8 M urea (Sigma), 3 M thiourea (Sigma), and β-mercaptoethanol for 30 min at room temperature prior to analysis. Total protein was resolved by Mini-PROTEAN GTX polyacrylamide gel electrophoresis (Bio-rad). Proteins were detected using anti-HA (Covance cat# MMS-101P, 1:1000) and anti-Actin (BD cat# 612657, 1:1000) antibodies. Blots were visualized using film or C-DiGit chemiluminescent imager (LI-COR).
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