Antibody was affinity purified using an epitope specific to EWS immobilized on solid support. The epitope recognized by A300-418A maps to a region between residue 600 and the C-terminus (residue 656) of human Ewing sarcoma breakpoint region 1 using the numbering given in SwissProt entry Q01844 (GeneID 2130). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
bK984G1.4, Ewing sarcoma breakpoint region 1, Ewing sarcoma breakpoint region 1 protein, Ewings sarcoma EWS-Fli1 (type 1) oncogene, EWS, EWS oncogene, EWS RNA-binding protein variant 6, EWS-FLI1, RNA-binding protein EWS
between 600 and C-term
2 - 8°C
Detection of human and mouse EWS by western blot. Samples: Whole cell lysate (15 and 50 µg) from HeLa and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-EWS antibody A300-418A (lot A300-418A-2) used for WB at 0.04 µg/ml. Detection: Chemiluminescence with an exposure time of 3 seconds.
Detection of human EWS 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-EWS antibody A300-418A (lot A300-418A-2) used for IP at 6 µg per reaction. EWS was also immunoprecipitated by rabbit anti-EWS antibody A300-417A. For blotting immunoprecipitated EWS, A300-418A was used at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 seconds.