Dig alkaline phosphatase fragment antigen binding fragment

The localization of LsHSCARB mRNA was detected in the adult female salmon louse by using in situ hybridization (ISH) as earlier described^{57 (link),58 (link)}. An RNA antisense (AS) probe of 731 bp was made from a target specific cDNA template (see Table 1 for primer sequences). A sense (S) probe acted as negative control for the transcript localization, whereas hybridization with a known set of probes detecting LsTryp1 acted as positive control^{59 (link)}. The labelled probes were visualized by using anti-digoxigenin (DIG) alkaline phosphatase fragment antigen binding (FAB) fragment (Roche) and a chromogen substrate containing levamisole (Sigma), nitroblue tetrazolium (NTB; Roche) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP; Roche). Microscopy images were captured by an Axio Scope A1 light microscope connected to Axiocam 105 (Zeiss).

The localization of LsRXR mRNA in adult female salmon lice was determined by in situ hybridization as described previously [20 (link)]. One 681bp and one 508bp RNA probe was synthesized on cDNA using the primers listed in Table 5. Hybridizations were performed with antisense probes to localize transcripts and sense probes as a negative control. Labeled probes were visualized using anti-digoxigenin (DIG) alkaline phosphatase fragment antigen binding (FAB) fragment (Roche) and a chromogenic substrate containing levamisol (Sigma–Aldrich), nitroblue tetrazolium (NTB; Roche) and 5-bromo-4-chloro-3-indolyl phosphate (BCIP; Roche).