Viewrna ez detection kit

ISH staining was performed on 4 μm-thick FFPE sections of six LGCA and six HGCA tissue samples and their patient-matched NC tissue samples. This was carried out on the Leica BOND^™ RX Auto-stainer using probes for OCT4 (NM_002701), SOX2 (NM_003106), NANOG (NM_024865), KLF4 (NM_004235) and c-MYC (NM_002467), using the ViewRNA eZ Detection Kit to detect the presence of mRNA (Affymetrix, Santa Clara, CA, USA). Positive controls were human seminoma for OCT4, NANOG and KLF4, normal skin for SOX2, and normal colon tissue for c-MYC. To determine specificity of the probes, negative controls were run using a probe for Bacillus (NM_L38424).

The 4μm-thick formalin-fixed paraffin-embedded CAML sections of six patients from the original cohort of nine patients used for DAB IHC staining were used for CISH staining using the Leica Bond Rx autostainer and detected using the ViewRNA eZ Detection kit (Affymetrix, CA, USA) as previously described (27 (link)). Probes for the stem cell markers c-Myc (NM_002467), KLF4 (NM_004235), NANOG (NM_024865), SOX2 (NM_003106), and OCT4 (POU5F1, NM_002701) were obtained from Affymetrix. Positive controls were demonstrated on human tissues: breast cancer for KLF4, normal prostate for c-Myc, skin for SOX2, and seminoma for OCT4 and NANOG. Negative control was demonstrated on a sample of Bacillus (NM_L38424).