Western Blot Analysis of Bacterial Proteins
Corresponding Organization : University of California, Santa Cruz
Other organizations : University of Wisconsin–Madison, University of South Carolina
Variable analysis
- Presence of dithiothreitol (DTT) in the final sample buffer
- Protein expression levels of IscR, YopD, YopE, and RpoA
- Number of cells used for protein sample normalization, as measured by OD600
- Sodium dodecyl sulfate-polyacrylamide gel (SDS-PAGE) used for protein separation
- Wet mini trans-blot cell (Bio-Rad) used for protein transfer to Immobilon-P membrane
- Blocking solution (Tris-buffered saline with Tween 20 and 5% skim milk)
- Primary antibodies (rabbit anti-IscR, rabbit anti-YopD, goat anti-YopE, rabbit anti-RpoA)
- Horseradish peroxidase-conjugated secondary antibodies
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
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