Rabbit anti zo 1 antibody

KW3110 was prepared as described previously [50 (link)]. Four strains of LAB other than KW3110 were prepared by isolation and culture from Japanese major commercial yogurt products, named strains A–D. LPS was purchased from Invivogen (San Diego, CA, USA). ATP was from Sigma (St. Louis, MO, USA). Hoechst 33342 and PI were from Dojindo (Tokyo, Japan). Rabbit anti-Claudin-1 antibody, rabbit anti-ZO-1 antibody, and goat anti-rabbit Immunoglobulin G (IgG) H&L (Alexa Fluor 488) were from Abcam (Cambridge, UK).

The distal portion of the colons from mice maintained on custom diets as well as those treated with CAY10397 or 15d-PGJ₂ and their respective vehicle controls were stored in 10% (v/v) buffered formalin. The sections were prepared at the Histopathology Core Facility, Animal Diagnostic Laboratory at Penn State University, University Park, PA. To detect Zo-1, sections were deparaffinized, followed by antigen-retrieval, and blocking with 5% goat serum. The sections were incubated in rabbit anti-Zo-1 antibody (1:500; Abcam) overnight at 4°C. The sections were washed in TBS and 0.5% hydrogen peroxide used to block any endogenous peroxidase. The sections were incubated in goat anti-rabbit secondary antibody (Vector Laboratories) for 1 h followed by incubation in ABC reagent (Vector Laboratories) for 45 min at room temperature. DAB substrate kit (Vector Laboratories) was used to develop peroxidase activity and the sections were dehydrated and mounted in dibutylphthalate polystyrene xylene (Sigma). The slides were imaged using an OLYMPUS DP74 microscope and OLYMPUS CellSens Standard software at magnification 10x. The length of well-oriented crypts of the distal colon of individual mice was also measured using the OLYMPUS CellSens Standard software.