Antidigoxigenin ap conjugate antibody

DNA probes were labelled with digoxigenin using the DIG-High Prime kit (Roche). Membranes were pre-hybridized in a 20 ml pre-hybridization solution (2x SSPE, 0.5% Blotto, 1% SDS, 10% dextran sulphate and 0.5 mg/ml⁻¹ sonicated and denatured salmon sperm DNA) at 65°C for 4–6 h. Labeled DNA was added and hybridization lasted for 12–16 h. Hybridized membranes were sequentially washed with 2x SSC and 0.1% SDS at room temperature for 5 min twice and with 0.1x SSC and 0.1% SDS at 68°C for 15 min twice as well. Detection was performed with an antidigoxigenin-AP conjugate antibody (Roche) and CDP-Star (Perkin Elmer) according to the instructions provided by the manufacturer. Quantification of autoradiograms was performed using a ImageJ64 software.

DNA probes were labeled with digoxigenin using the DIG-High Prime kit (Roche). Membranes were prehybridized in a 20 ml prehybridization solution (2× SSPE (Saline Sodium-Phosphate-EDTA), 0.5% Blotto, 1% SDS, 10% dextran sulfate and 0.5 mg/ml sonicated and denatured salmon sperm DNA) at 65ºC for 4–6 h. Labeled DNA was added and hybridization lasted for 12–16 h. Hybridized membranes were sequentially washed with 2× SSC (Saline Sodium-Citrate) and 0.1% SDS at room temperature for 5 min twice and with 0.1× SSC and 0.1% sodium dodecyl sulfate (SDS) at 68ºC for 15 min twice as well. Detection was performed with an antidigoxigenin-AP conjugate antibody (Roche) and CDP-Star (Perkin Elmer) according to the instructions provided by the manufacturer.