Antibody was affinity purified using an epitope specific to PBEF immobilized on solid support. The epitope recognized by A300-372A maps to a region between residues 400 and 450 of human pre-B-cell colony enhancing factor 1 using the numbering given in Swiss-Prot entry P43490 (GeneID 10135). Immunoglobulin concentration was determined by extinction coefficient: absorbance at 280 nm of 1.4 equals 1.0 mg of IgG.
1110035O14Rik, NAmPRTase, Nampt, nicotinamide phosphoribosyltransferase, PBEF, PBEF1, pre-B cell-enhancing factor, pre-B-cell colony-enhancing factor 1, VF, VISFATIN
between 400 and 450
2 - 8°C
Detection of human and mouse PBEF 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-PBEF antibody A300-372A (lot A300-372A-5) used for WB at 0.1 µg/ml. Detection: Chemiluminescence with an exposure time of 10 seconds.
Detection of human PBEF 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-PBEF antibody A300-372A (lot A300-372A-5) used for IP at 6 µg per reaction. PBEF was also immunoprecipitated by rabbit anti-PBEF antibodies A300-778A and A300-779A. For blotting immunoprecipitated PBEF, A300-372A was used at 1 µg/ml. Detection: Chemiluminescence with an exposure time of 30 seconds.