Antibody was affinity purified using an epitope specific to Cul7 immobilized on solid support. The epitope recognized by A300-223A maps to a region between residues 1500 and 1550 of human Cullin homolog 7 using the numbering given in SwissProt entry Q14999 (GeneID 9820). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
3M1, CUL-7, cullin-7, dJ20C7.5, KIAA0076
between 1500 and 1550
2 - 8°C
Tris-citrate/phosphate buffer, pH 7 to 8 containing 0.09% Sodium Azide
Detection of human and mouse Cul7 by western blot. Samples: Whole cell lysate (50 µg) from Hep-G2, HEK293T, HeLa, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-Cul7 antibody A300-223A (lot A300-223A-4) used for WB at 0.4 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.
Detection of human Cul7 by western blot of immunoprecipitates. Samples: Whole cell lysate (1.0 mg per IP reaction; 20% of IP loaded) from Hep-G2 cells prepared using NETN lysis buffer. Antibodies: Affinity purified rabbit anti-Cul7 antibody A300-223A (lot A300-223A-4) used for IP at 6 µg per reaction. Cul7 was also immunoprecipitated by a previous lot of this antibody (lot A300-223A-3) and rabbit anti-Cul7 antibody A300-224A. For blotting immunoprecipitated Cul7, A300-223A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 75 seconds.