Cd34 antibody clone

For 35 cases we performed CD34 immunostaining. The tissue samples were derived from formalin-fixed paraffin-embedded tissue blocks with primary CRC and tumor-free margin specimens. Then, samples underwent deparaffinization and rehydration. In the next step, antigen retrieval was performed by cooking slides in EnVision Flex Target Retrieval Solution High pH (Dako, Carpinteria, CA, USA) for 20 min at 95 °C. Prepared samples were incubated with a peroxidase-blocking reagent (Dako) and then incubated with CD34 antibody (clone: QBEnd/10 Cell Marque, Rocklin, CA, USA; incubation time: 30′; dilution: 1:150). Subsequently, they were put in EnVision FLEX HRP (Dako). Next, antigen–antibody complexes were stained using 3.3′-diaminobenzidine. Finally, tissue sections were counterstained with hematoxylin, dehydrated, and covered with coverslips for further analysis.

CD 34 immunostaining was performed in 24 randomly selected cases. Tissue samples were obtained from formalin-fixed paraffin-embedded tissue blocks with primary CRC and tumor-free margin samples. Then, the samples were deparaffinized and rehydrated. We then performed antigen recovery by boiling the slides in EnVision Flex Target Retrieval Solution High pH (Dako, Carpinteria, CA, USA) for 20 min at 95 °C. Prepared samples were incubated with peroxidase blocked reagent (Dako, Carpinteria, CA, USA) and then incubated with CD34 antibody (clone: QBEnd/10 Cell Marque, Rocklin, CA, USA; incubation time: 30′; dilution: 1:150). In the next step, the samples were placed in EnVision FLEX HRP (Dako, Carpinteria, CA, USA). The antigen-antibody complexes were then stained with 3,3′-diaminobenzidine. Finally, tissue sections were counterstained with hematoxylin, dehydrated and covered with coverslips for further analysis.