ANTIBODIES TO NUCLEOPORIN PROTEINS

The nuclear pore complex (NPC) is a structure that spans the nuclear envelope and provides a conduit between the nucleus and cytoplasm for the nucleocytoplasmic transport of macromolecules. The main components of the NPC are nucleoporins. There are about 30 different nucleoporin proteins that have been identified and classified into 3 families: transmembrane nucleoporins, FG- or FN-nucleoporins, and WD-repeat seven-bladed propeller motif nucleoporins. Nucleoporins exists in a wide range of sizes and are named according to their molecular weight in kilodaltons. Nucleoporins appear to serve non-redundant functions in the NPC, however their roles in the NPC are not fully understood. Some have been shown to participate as essential structural elements that assemble into subcomplexes for the formation of an NPC with eight-fold symmetry. Others have been suggested to play important roles in the active transport of macromolecules across the NPC; however none have been identified that exhibit motor or ATPase/GTPase activities. Recent studies of leukemogenic fusion proteins that contain functional elements of nucleoporins have brought to light a possible novel function of nucleoporins in transcription. These findings illustrate that the functional roles of nucleoporins are likely to be expansive and varied, and much remains to be learned about the roles of the nucleoporins in NPC assembly, nucleocytoplasmic transport, and other cellular processes. 

Detection of Human NUP50 by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 mcg for Western Blot; 1 mg for immunoprecipitation, 20% of IP loaded) from HeLa cells. Antibody: Affinity purified rabbit anti-NUP50 antibody (Cat. No. A301-782A) used for WB at 0.1 mcg/ml (A) and 1 mcg/ml (B) and used for IP at 3 mcg/mg lysate. NUP50 was also mmunoprecipitated by rabbit anti-NUP50 antibody (Cat. No. A301-783A), which recognizes a downstream epitope. For blotting immunoprecipitated NUP50, the ReliaBLOT® Reagents and Procedures (Cat. No. WB120) were used. Detection: Chemiluminescence with exposure times of 30 seconds (A) and 3 seconds (B).

Detection of Human NUPL by Western Blot and Immunoprecipitation. Samples: Whole cell lysate from HeLa (5, 15 and 50 mcg for Western Blot; 1 mg for Immunoprecipitation, 20% of IP loaded) and 293T (T; 50 mcg) cells. Antibody: Affinity purified rabbit anti-NUPL antibody (Cat. No. A301-792A) used for Western Blot at 0.04 mcg/ml (A) and 1 mcg/ml (B) and used for Immunoprecipitation at 3 mcg/mg lysate. NUPL was also immunoprecipitated by a rabbit anti-NUPL antibody, which recognizes an upstream epitope. For blotting immunoprecipitated NUPL, the ReliaBLOT® Reagents and Procedures (Cat. No. WB120) were used. Detection: Chemiluminescence with exposure times of 10 seconds (A) and 3 seconds (B).

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