Anti-CRISP3 antibody (ab105951)

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Freshly cut tissue microarray sections were analyzed in 1 day and in one experiment. A polyclonal CRISP3 antibody (rabbit, Abcam; at 1/450 dilution; cat#ab105951, Cambridge, UK) was used for detection of CRISP3 expression. Slides were deparaffinized and exposed to heat-induced antigen retrieval for 5 min in an autoclave at 121 °C in pH 7.8 buffer. Bound primary antibody was visualized using the DAKO EnVision Kit (Dako, Glostrup, Denmark). CRISP3 staining was evaluated according to the following scoring system as previously described:21, 22, 23 The staining intensity (0, 1+, 2+, and 3+) and the fraction of positive tumor cells were recorded for each tissue spot. A final score was built from these two parameters as follows: Negative: absence of CRISP3 staining; weak: intensity of 1+ in ≤70% of tumor cells or staining intensity of 2+ in ≤30% of tumor cells; moderate: intensity of 1+ in >70% of tumor cells, or staining intensity of 2+ in >30% but ≤70% of tumor cells or staining intensity of 3+ ≤30% of tumor cells; strong: intensity of 2+ in >70% of tumor cells or staining intensity of 3+ in >30% of tumor cells.

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