Ventana universal secondary antibody

Monoclonal antibodies (mAb) for Ki67 (8D5) and psoriasin (47C1068) were purchased from Cell Signaling Tech (Danvers, MA 01923) and Abcam (Cambridge, UK), respectively. The non-steroidal anti-inflammatory (NSAID), ibuprofen (IB), was obtained from Sigma–Aldrich (St. Louis, MO, USA). IHC staining of reconstituted psoriatic and normal human tissue models was conducted. Deparaffinized sections were subjected to antigen retrieval, followed by incubation with primary antibodies (1:500 dilution for Ki67 mAb and 1:200 dilution for anti-psoriasin mAb). Bound antibodies were detected by DAB staining using Ventana universal secondary antibody (Ventana Medical System, Tuscan, Arizona). All stained slides were digitally imaged at magnification equivalent to ×20. Representative fields were scored by an independent pathologist.

NF-κB p-p65 monoclonal antibody (93H1) was from Cell Signaling Tech (Danvers, MA, United States), and PDE4 polyclonal antibody (ab14628) was from Abcam (Cambridge, United Kingdom). The BIOMOL Cyclic Nucleotide PDE Assay kit was from ENZO Life Sciences, Inc. (Farmingdale, NY, United States). LPS (Escherichia coli 0111:B4) and 3-isobutyl-1-methylxanthine (IBMX) were from Sigma-Aldrich (St. Louis, MO, United States), and rolipram was a gift from MedChem Express (Monmouth Junction, NJ, United States).
Immunohistochemistry staining of reconstituted psoriatic and normal tissue models was conducted using deparaffinized sections subjected to antigen retrieval, followed by incubation with anti-PDE4 primary antibody (1:100). Bound antibody was detected by DAB staining using a Ventana universal secondary antibody (Ventana Medical System, Tuscan, AZ, United States). All stained slides were digitally imaged at magnification equivalent to 20×. Random fields were scored by an independent pathologist.