Frozen tissues were pulverized prior to protein extraction essentially as above. Total RNA was extracted using the Direct-zol RNA isolation kit (Genessee Scientific) per manufacturer's instructions. mRNA expression was detected as above. IHC analysis was conducted as described (13 (link)). Using antibodies purchased from Thermo Scientific (ESR1, RM-9101-S), (Ki67, RM-9106-S1) and Cell Signaling (pRb S807/811, 8516).
Protocol detail
Tissue Pulverization and RNA/Protein Analysis
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Corresponding Organization : Washington University in St. Louis
Other organizations : Baylor College of Medicine, Pfizer (United States), Saint Louis University
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Variable analysis
independent variables
- Pulverized frozen tissues
- MRNA expression
- Protein expression (detected by IHC analysis)
- Extraction protocols (Direct-zol RNA isolation kit, manufacturer's instructions)
- Antibodies used for IHC analysis (ESR1, Ki67, pRb S807/811)
- Positive control: Not specified
- Negative control: Not specified
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