Cell Signaling Technology

Product Pathways - Cytoskeletal Signaling

Phospho-RanBP3 (Ser58) Antibody #9380

Applications Reactivity Sensitivity MW (kDa) Source
W IP H Mk Endogenous 70 kDa Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-RanBP3 (Ser58) Antibody recognizes endogenous levels of RanBP3b protein only when phosphorylated at Ser58, and RanBP3a protein only when phosphorylated at corresponding residue Ser126.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser58 of human RanBP3b protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa and COS-7 cells, serum-starved overnight and either left untreated or treated with hEGF #8916 (100 ng/ml), using Phospho-RanBP3 (Ser58) Antibody (upper) or RanBP3 Rabbit mAb (lower).

Background

RanBP3 was originally identified as RanGTP binding protein located in the nucleus and involved in the nuclear exporting process (1). It functions as a cofactor for CRM1 nuclear export by binding to CRM1, stabilizing the RanGTP-CRM1-cargo interaction and promoting complex association with nuclear pore proteins (2,3). In the absence of Ran-bound GTP, RanBP3 prevents binding of CRM1 complex to the nuclear pore complex. In addition to CRM1, RanBP3 also has been shown to bind to RanGEF-RCC1 and increase the guanine nucleotide exchange activity of RCC1 for RanGTP-CRM1-Cargo (1,4). In some cases, as with β-catenin and Smad2/3, RanBP3 binding may mediate the target protein nuclear export in a Ran-dependent, but CRM1-independent manner (5,6). RanBP3 is phosphorylated at Ser58 through the PI3K/Akt or ERK/RSK pathway. This phosphorylation is important for RanBP3 function in nuclear export, likely due to stimulation of RCC1 activity (7,8).

  1. Mueller, L. et al. (1998) FEBS Lett 427, 330-6.
  2. Lindsay, M.E. et al. (2001) J Cell Biol 153, 1391-402.
  3. Englmeier, L. et al. (2001) EMBO Rep 2, 926-32.
  4. Nemergut, M.E. et al. (2002) J Biol Chem 277, 17385-8.
  5. Hendriksen, J. et al. (2005) J Cell Biol 171, 785-97.
  6. Dai, F. et al. (2009) Dev Cell 16, 345-57.
  7. Yoon, S.O. et al. (2008) Mol Cell 29, 362-75.
  8. von Knethen, A. et al. (2010) J Cell Sci 123, 192-201.

Application References

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