MyD88 and Smad7 expression were determined by Western blot using antibodies to MyD88 (1:200) or Smad7 (1:300; Santa Cruz Biotechnology) using our previously-described protocol25 (link). Blots were probed with horseradish peroxidase-conjugated secondary antibodies 1:5,000–1:10,000 in blocking buffer for 1 h, and bands were detected using the chemiluminescent method (SuperSignal West Dura, Thermo Scientific, Waltham, MA). Blots were stripped and re-probed with anti-actin (Santa Cruz Biotechnology) to ensure equal loading.
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Western Blot Analysis of MyD88 and Smad7
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Corresponding Organization : Birmingham VA Medical Center
Other organizations : Nationwide Children's Hospital
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Variable analysis
independent variables
- MyD88 expression
- Smad7 expression
- MyD88 protein levels
- Smad7 protein levels
- Equal loading of samples, as ensured by re-probing the blots with anti-actin antibody
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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