Antibody was affinity purified using an epitope specific to Fen1 immobilized on solid support. The epitope recognized by A300-255A maps to a region between residues 75 and 125 of human Flap Structure Endonuclease 1 using the numbering given in entry NP_004102.1 (GeneID 2237). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
DNase IV, FEN-1, flap endonuclease 1, Flap structure-specific endonuclease 1, hFEN-1, Maturation factor 1, maturation factor-1, MF1, RAD2
between 75 and 125
2 - 8°C
Detection of human and mouse Fen1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa, HEK293T, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-Fen1 antibody A300-255A (lot A300-255A-3) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human Fen1 by western blot of immunoprecipitates. Samples: Whole cell lysate (0.5 or 1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Fen1 antibody A300-255A (lot A300-255A-3) used for IP at 6 µg per reaction. Fen1 was also immunoprecipitated by rabbit anti-Fen1 antibody A300-256A. For blotting immunoprecipitated Fen1, A300-255A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.