Antibody was affinity purified using an epitope specific to INT1 immobilized on solid support. The epitope recognized by A300-361A maps to a region between residues 150 and 200 of human integrator complex subunit 1 using the numbering given in entry NP_001073922.1 (GeneID 26173). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
INT1, integrator complex subunit 1, NET28
between 150 and 200
2 - 8°C
Tris-citrate/phosphate buffer, pH 7 to 8 containing 0.09% Sodium Azide
Detection of human and mouse INT1 by western blot. Samples: Whole cell lysate (50 µg) from HeLa, HEK293T, Jurkat, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-INT1 antibody A300-361A (lot A300-361A-2) used for WB at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human INT1 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from HeLa cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-INT1 antibody A300-361A (lot A300-361A-2) used for IP at 3 µg per reaction. INT1 was also immunoprecipitated by rabbit anti-INT1 antibody BL998. For blotting immunoprecipitated INT1, A300-361A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 3 minutes.