Dako target retrieval solution ph9

Skin and tumors were fixed in 10% buffered formalin and embedded in paraffin. Sections were stained with H&E and histopathological evaluation was performed by two experimented observers: MJFA, specialized in human pathology and RAGF, a veterinarian expert in animal pathology.
Immunostaining was performed using antibodies against IKKα (NB100-56704) IKKβ (Novus Biologicals, Cambridge UK); IKKα (H00001147-M04) (Abnova, Taiwan); IKKα (sc-7182), P-IKKα/β (Ser 180/Ser 181)-R (sc-23470-R), Maspin, p65 (Santa Cruz Biotechnology, Inc. Heidelberg, Germany); CD31, E- Cadherin, Integrin-α6 (BD Bioscience, NJ, USA); p52 (Abcam, Cambridge, UK). Sections were incubated with a biotinylated secondary antibody, and then with streptavidin conjugated to horseradish peroxidase (DAKO A/S, Glostrp, Denmark). Antibody localization was determined using 3,3-diaminobenzidine (DAB) (Vector Laboratories; Burlingame, CA, USA).
A pressure cooker with DAKO target retrieval solution ph9.0 (DAKO) was employed for Maspin, mouse IKKα, human IKKα, P-IKKα/β, IKKβ and E- Cadherin detection. Staining with p52, p65, Integrin-α6 and CD31 antibodies was performed in cryosections of tumors.

Skin equivalents were fixed in 10% buffered formalin and embedded in paraffin. Sections were stained with H&E and histopathological evaluation of skin equivalents was performed by two specialists in pathological anatomy: MJFA, specialized in human pathology and AB, a veterinarian expert in animal pathology. Immunostaining was performed using antibodies against IKKα NB-100-56704 (Novus Biologicals, Cambridge UK); IKKα sc-7182 (Santa Cruz Biotechnology, Inc. Heidelberg, Germany); IKKα 556532, Plakoglobin (BD Bioscience, NJ, USA); Involucrin, Filaggrin (Covance, CA, USA); BrdU (Roche, Mannheim, Germany). Sections were incubated with a biotinylated secondary antibody, and then with streptavidin conjugated to horseradish peroxidase (DAKO A/S, Glostrp, Denmark). Antibody localization was determined using 3,3-diaminobenzidine (DAB) in H₂O (Vector Laboratories; Burlingame, CA, USA).
Pressure cooker with DAKO target retrieval solution ph9.0 (DAKO) was employed for antibodies detection.

FFPE tissue sections (3.5 μm) were deparaffinized and rehydrated for immunohistochemical staining. Slides with tissues sections were incubated for heat‐induced antigen retrieval in Dako Target Retrieval Solution Citrate pH 6.0 (Dako S2369) or Dako Target Retrieval Solution pH 9,0 (Dako S2367) for 30 minutes in a steamer. The staining was then performed manually at 4°C antibody incubation using the Dako REAL™ Detection System, Peroxidase/AEC, using monoclonal antibodies directed against: HBD2 (1:400; #ab63982, Abcam Cambridge, U.K.), HBD3 (1:100; #LS‐B86, LSbio Seattle, WA), psoriasin (1:300; #MA1‐91555, Thermo Fisher Scientific, Pittsburgh, PA), RNase7 (1:50; #ab154143, Abcam Cambridge, U.K.) and LL37 (1:50; #63982, Abcam Cambridge, U.K.). Images of stainings were acquired with a DP71 digital camera (Olympus, Vienna, Austria), attached to an Olympus BX51 microscope.