Tgf β primary antibody

The sections were stained with hematoxylin-eosin (HE) and Picrosirius Red for histological evaluation and fixed in 10% paraformaldehyde, and embedded in paraffin. Then, 5-μm-thick sections were cut from the blocks, affixed to glass slides coated with poly-L-lysine, examined under a microscope (Nikon Eclipse E400; Nikon, Tokyo, Japan), and evaluated with a digital image analysis system. Immunohistochemically analysis was performed as previously described by Jones et al. 2008 [18 (link)]. Briefly, the skin samples were incubated with a primary antibody applied for 1 h at room temperature. Tissue sections were then incubated with TGF-β primary antibody (1:200, Abcam Inc; http://www.abcam.com, accessed on 28 February 2023) for 1 h at 25 °C and incubated with primary antibodies overnight at 4 °C, followed by incubation with secondary antibodies for 2 h at 25 °C, followed by washing and visualized using an isothiocyanate-conjugated secondary antibody (Jackson Immuno Research Laboratories Inc., West Grove, PA, USA) and imaged with an optical microscope Axio Scan.Z1 scanner (Carl Zeiss, Germany).

After scanning, the specimens were decalcified, dewatered, and embedded in paraffin. Then, they were mounted in the medial plane on 4-μm serial sections on anti-adhesive slides in preparation for HE staining, Masson staining, as well as immunohistochemical (IHC) staining. IL-1β primary antibody (dilution 1:1000, Abcam, USA) and TGF-β primary antibody (dilution 1:1000, Abcam, USA) was used for IHC staining. Three consecutive square fields involving junctional epithelium (JE), the gingival epithelium (GE), and alveolar bone (AB) in the mesial coronal area of the first molar were selected from each section. Six sections of each group were analyzed, and each measurement was counted by the examiner in a blind manner.