Antibody was affinity purified using an epitope specific to 4E-t/eIF4E-T immobilized on solid support. The epitope recognized by A300-706A maps to a region between residue 925 and the C-terminus (residue 985) of human Eukaryotic Translation Initiation Factor 4E Nuclear Import Factor 1 using the numbering given in entry NP_062817.1 (GeneID 56478). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
2610509L04Rik, 4E-T, Clast4, eIF4E transporter, Eukaryotic translation initiation factor 4E nuclear import factor 1, eukaryotic translation initiation factor 4E transporter
between 925 and C-term
2 - 8°C
Detection of human and mouse 4E-T/eIF4E-T 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-4E-T/eIF4E-T antibody A300-706A (lot A300-706A-2) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human 4E-T/eIF4E-T 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-4E-T/eIF4E-T antibody A300-706A (lot A300-706A-2) used for IP at 6 µg per reaction. 4E-T/eIF4E-T was also immunoprecipitated by a previous lot of this antibody A300-706A-1. For blotting immunoprecipitated 4E-T/eIF4E-T, A300-706A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human 4E-T by immunohistochemistry. Sample: FFPE section of human colon carcinoma. Antibody: Affinity purified rabbit anti-4E-T (Cat. No. A300-706A Lot2) used at a dilution of 1:1,000 ( 1µg/ml). Detection: DAB