Fitc conjugated donkey anti rabbit igg antibody

All skin sections were taken from the para-midline, lower-back region. Sections were stained with hematoxylin and eosin. We examined dermal thickness, which was defined as the thickness of skin from the dermal-epidermal junction to the junction between the dermis and subcutaneous fat. The thickness of dermis and subcutaneous fat was measured from seven different randomly selected fields per specimen. Immunohistochemistry was performed using antibodies directed at α-SMA (Sigma-Aldrich, St. Louis, MO), Smad3 (Santa Cruz Biotechnology, Santa Cruz, CA), and arginase-1 (Santa Cruz Biotechnology). For immunofluorescence, goat anti-VE-cadherin antibody (Santa Cruz Biotechnology) and rabbit anti-FSP-1 antibody (Abcam, Cambridge, UK) were used as primary antibodies, and FITC-conjugated donkey anti-rabbit IgG antibody (Santa Cruz Biotechnology) and Alexa Fluor donkey 555 anti-goat IgG antibody (Invitrogen, Carlsbad, CA) were used as secondary antibodies. Coverslips were mounted by using Vectashield with DAPI (Vector Laboratories, Burlingame, CA), and staining was examined by using Bio Zero BZ-8000 (Keyence, Osaka, Japan) at 495 nm (green), 565 nm (red), and 400 nm (blue). All sections were examined independently by two investigators in a blinded manner.