Journal: Experimental dermatology

Article Title: On the relationship between VDR, RORα and RORγ receptors expression and HIF1-α levels in human melanomas

doi: 10.1111/exd.14002

Figure Lengend Snippet: Characteristic of primary and secondary antibodies and visualization systems used in IHC staining.

Article Snippet: For each immunostaining run also appropriate positive controls were prepared ( ). table ft1 table-wrap mode="anchored" t5 Table 1. caption a7 Antibody (vendor) Primary antibody dilution * Secondary antibody (vendor) Chromogen (vendor) Positive control Mouse anti-HIF-1α (Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat# sc-53546) 1:750 ** EnVision+System HRP Labelled Polymer (Dako, Glostrup, Denmark, Cat# K4000) Vector NovaRED HRP substrate (Vector Laboratories, Inc., Burlingame, CA, USA, Cat#SK-4800) breast cancer Rat anti-VDR (Abcam, Cambridge, MA, Cat#ab8756) 1:75 Vectastain ABC-AP Kit (Vector Laboratories Inc, Burlingame, CA, Cat#AK-5004) Vectastain ABC-AP Reagent and alkaline phosphatase substrate (Vector Laboratories Inc, Burlingame, CA, USA, and SK5100). normal skin/kidney Rabbit anti-CYP27B1 (Santa Cruz Biotechnology, Santa Cruz, CA, Cat#sc-67261) 1:75 EnVision+System HRP Labelled Polymer (Dako, Glostrup, Denmark, Cat# K4002) ImmPACT NovaRED (Vector Laboratories Inc., Burlingame, CA, USA, Cat#SK-4805) kidney Rabbit anti-CYP24A1 (Abcam, Cambridge, MA) 1:40 EnVision+System HRP Labelled Polymer (Dako, Glostrup, Denmark) ImmPACT NovaRED (Vector Laboratories Inc., Burlingame, CA, USA) kidney Rabbit anti-RORγ (Dr Anton M. Jetten) 1:50 EnVision™ FLEX/HRP (Dako, Glostrup, Denmark, Cat# K4002) Vector NovaRED HRP substrate (Vector Laboratories, Inc., Burlingame, CA, USA, Cat#SK-4800) normal skin Goat-antiROR α (Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat#sc-6062) 1:25 Vectastain Elite ABC Goat IgG (Vector Laboratories, Burlingame, CA, USA, Cat#PK6105) ImmPACT™ NovaRED™ HRP substrate (Vector Laboratories, Cat#SK-4805) normal skin Mouse anti-FoxP3 (Abcam, Cambridge, UK, Cat#ab20034) 1:100 EnVision+System HRP Labelled Polymer (Dako, Glostrup, Denmark, Cat# K4000) Vector NovaRED HRP substrate (Vector Laboratories, Inc., Burlingame, CA, USA, Cat#SK-4800) lymph node Open in a separate window ON – overnight; * primary antibodies were applied overnight at 4°C; ** Tyramide Signal Amplification kit (Life Technologies, Carlsbad, CA, Cat# {"type":"entrez-nucleotide","attrs":{"text":"T20931","term_id":"2756849","term_text":"T20931"}} T20931 ) was used according the manufacturer’s protocol Characteristic of primary and secondary antibodies and visualization systems used in IHC staining.

Techniques: Staining, Positive Control, Plasmid Preparation