Fluorescein isothiocyanate fitc conjugated mouse anti digoxygenin antibodies

After SC analysis, the same cells were subjected to FISH with BAC clones (SBAB-428G6, SBAB-413G8, SBAB-498D8 from the National Institute for Agricultural Research (INRA) BAC library) (Rogel-Gaillard et al. 1999 (link)): one was located in the telomeric region of the SSC1 p arm (labeled with biotin), and two were located on SSC14, one in the telomeric region (labeled with digoxigenin) and the other in the centromeric region (containing porcine endogenous retrovirus (PERV) sequences, differentially labeled with biotin and digoxigenin). These BAC clones were labeled using the BioPrime DNA Labeling System kit (Life Technologies, Carlsbad, CA, USA), and revealed by Alexa 594 conjugated to streptavidin (Molecular Probes, Eugene, OR, USA) and fluorescein isothiocyanate (FITC) conjugated mouse anti-digoxygenin antibodies (Sigma-Aldrich, Saint Louis, MO, USA). FISH signals from the same cells for which SCs had previously been analyzed were captured and evaluated using an Imager Z2 microscope with CytoVision imaging system (Leica Microsystemes, Nanterre, France).

After SC analysis, the same cells were subjected to fluorescence in situ hybridization with BAC clones: one was located in the telomeric region of the SSC1 p-arm (labeled with biotin), and one in the centromeric region of SSC15. These BAC clones were labeled using the BioPrime DNA Labeling System kit (LIFE TECHNOLOGIES, Carlsbad, CA, USA), and revealed by Alexa 594 conjugated to Streptavidin (MOLECULAR PROBES, Eugene, OR, USA) and fluorescein isothiocyanate (FITC) conjugated mouse anti-digoxygenin antibodies (SIGMA-ALDRICH, St. Louis, MO, USA). FISH signals from the same cells for which SCs had previously been analyzed were captured and evaluated using an Imager Z2 microscope with the Cytovision imaging system (LEICA MICROSYSTEMES, Nanterre, France).