Horseradish peroxidase conjugated rabbit igg

NSCLC cancerous and matched adjacent non-cancerous tissues were prepared for TMAs. The manual Tissue Microarray System Quick-Ray (UT06, UNITMA, Korea) was used to generate TMAs. Core tissue samples (diameter of 2 mm) were obtained from paraffin embedded tissue sections and deposited in paraffin-recipient blocks. Before immunohistochemical processing, the tissue microarray specimens were cut into 4-μm-thick sections and placed on the super frost charged glass microscope slides. IHC was carried out as described before [50 (link)]. Briefly, the slides were incubated with the primary antibodies for FABP3 (1:100; Abcam, Cambridge, MA, USA) and FABP4 (1:100; Abcam, Cambridge, MA, USA) overnight at 4°C. Subsequently, the slides were incubated with anti-mouse IgG secondary antibody (Abcam) for FABP3 and horseradish-peroxidase-conjugated rabbit IgG (Abcam) for FABP4. Diaminobenzidine solution was used for the color development.

NSCLC and matched, tumor-adjacent tissues were prepared and used for TMAs. The TMAs were assembled using a tissue arraying instrument (Quick-Ray, UT06; UNITMA, Korea). Core tissue samples (2 mm in diameter) were taken from individual paraffin-embedded sections and deposited in recipient paraffin blocks. TMA specimens were cut into 4-μm sections and put on the super frost-charged glass microscope slides before immunohistochemical processing. Immunohistochemical analysis was performed as previously described. The slides were incubated with the primary antibodies against ROR2 (1:100; LS-C99125, LifeSpan BioSciences, Seattle, WA, USA) or Wnt5a (1:200; Abcam, Cambridge, MA, USA) at 4°C overnight. Horseradish-peroxidase-conjugated rabbit IgG (Abcam) was applied as the secondary antibody for ROR2, and anti-mouse IgG (Abcam) was applied for Wnt5a. The binding of the primary antibody was detected using diaminobenzidine solution. Slide in which primary antibody was omitted was used a negative control, while a breast cancer sample known to be ROR2 positive was included as a positive control.