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. 2019 Jun 24;9:9142. doi: 10.1038/s41598-019-45161-8

Table 1.

Description of the antibodies used and their manufacturers, and of immunocytochemistry protocols and detection methods.

Antibody Vendor Permeabilization/blocking before primary antibodies application Primary antibody (dilution; incubation time) Secondary antibody (dilution; incubation time; vendor)
Anti-VDR, rat monoclonal, clone 9A7 Abcam (Abcam Inc., Cambridge, MA, USA) NA 1:50, O/N, 4 °C anti-rat, peroxidase-conjugated goat antibody (1:250, 30 min, Jackson ImmunoResearch Laboratories, Inc. West Grove, PA, USA);
Anti-CYP27B1, rabbit polyclonal, clone H-90 Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA) 0.2% Triton-X100, 15 min, RT 1:75, O/N, 4 °C EnVision System-HRP (RTU, 30 min, antibody with peroxidase-conjugated polymer backbone,Dako, Carpinteria, CA, USA)
Anti-CYP24A1, mouse monoclonal, Abcam (Abcam Inc., Cambridge, MA, USA) 0.2% Triton-X100 in 1% BSA, 25 min, RT 1:30, O/N, 4 °C EnVision System-HRP (RTU, 30 min, antibody with peroxidase-conjugated polymer backbone, Dako, Carpinteria, CA, USA)
Anti-RORα, goat polyclonal, clone C-16 Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA) 2% BSA, 20 min, RT 1:500, O/N, 4 °C EnVision System-HRP (RTU, 30 min, antibody with peroxidase-conjugated polymer backbone,Dako, Carpinteria, CA, USA)
Anti-RORγ, rabbit customer-made at NIEHS by Dr Jetten* 0.1% Triton-X100 in 3.5% BSA, 60 min, RT 1:50, O/N, 4 °C EnVision System-HRP (RTU, 30 min, antibody with peroxidase-conjugated polymer backbone,Dako, Carpinteria, CA, USA)

RT – room temperature, O/N, overnight; NA – not applied; BSA- bovine serum albumin; RTU – ready-to-use; *for detailed description see47,62. The melanin content was assessed in routine pathomorphological slides (stained with hematoxylin and eosin) as previously described62,79.