Primary tumors and metastatic tissues were resected, formalin fixed and/or decalcified (bone) and sent to the University of Colorado Denver Biorepository Core where tissues were paraffin-embedded. Tumors were sectioned and stained using immunohistochemistry performed as previously described [21 (link)]. Antibodies used were: MUC2 (1:10000 Abcam ab134119) and MCM2 (Cell Signaling, 4007). Images were captured using the Aperio Digital Pathology system (Leica Biosystems) and positive staining was quantified via Imagescope software (Leica). Mucicarmine staining was performed as previously reported [21 (link)] and used to assess total secreted mucins and quantify necrotic regions. For mucicarmine staining, the Aperio imaging system was used followed by quantitation using the ImageJ software (FIJI).
Protocol detail
Immunohistochemical Analysis of Tumor Tissues
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Antibodies
Bone
Formalin
Immunohistochemistry
Mcm2
Muc2
Mucicarmine
Mucins
Necrotic
Paraffin
Tissues
Tumors
Corresponding Organization :
Other organizations : University of Colorado Anschutz Medical Campus
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Variable analysis
independent variables
- Primary tumors and metastatic tissues resected
- Positive staining quantified via Imagescope software (Leica)
- Total secreted mucins quantified
- Necrotic regions quantified
- Tissues were formalin fixed and/or decalcified (bone)
- Tissues were paraffin-embedded
- Immunohistochemistry performed as previously described [21]
- Antibodies used: MUC2 (1:10000 Abcam ab134119) and MCM2 (Cell Signaling, 4007)
- Mucicarmine staining performed as previously reported [21]
- Aperio imaging system used for mucicarmine staining, followed by quantitation using the ImageJ software (FIJI)
- Not explicitly mentioned
- Not explicitly mentioned
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