Western blot analysis of Exportin 1 was performed by loading 25 ug of SK-BR-3 (lane 1), Hela (lane 2) and NIH-3T3 (lane 3) onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with an Exportin 1 polyclonal antibody (Product # PA1-083) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~130kDa.
|Tested species reactivity||Bovine, Human, Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to residues C R(1043) Q A D E E K H K R Q M S V P G S(1058) of human CRM1.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Immunoprecipitation (IP)||Assay dependent|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
PA1-083 detects CRM1 (chromosome region maintenance 1) from bovine retinal extract, human and mouse samples.
PA1-083 has been successfully used in Western blot and immunoprecipitation procedures. By Western blot, this antibody detects an ~135 kDa protein representing CRM1 from bovine retinal extract.
The PA1-083 immunogen is a synthetic peptide corresponding to residues C R(1043) Q A D E E K H K R Q M S V P G S(1058) of human CRM1. This sequence is 88% conserved in mouse, rat, and Xenopus laevis. The PA1-083 immunizing peptide (Cat. # PEP-162) is available for use in neutralization and control experiments.
Diffusion of metabolites and small non-nuclear molecules as well as active, mediated import of protein and export of protein and RNA through the nuclear envelope occurs through nuclear pore complexes or NPC and quote;s. NPC and quote;s contain up to 100 different polypeptides which have a combined mass of about 125 megadaltons. The channel available for passive transport through the NPC is about 9-10 nm in diameter while carrier mediated changes in the NPC result in a ~25 nm channel used for larger, actively transported molecules.
Active transport of molecules through the NPC is a signal-mediated process that is not well understood. Proteins to be exported from the nucleus to cytoplasm contain leucine-rich nuclear export signals (NESs). NESs are recognized by transport receptors belonging to the importin-beta family of proteins. It has been shown that NES export is mediated by CRM1 (chromosome region maintenance 1), an importin-beta family member that binds NESs, Ran:GTP, and Ran binding protein (RanBP). CRM1 mediates the nuclear exportation of numerous proteins, including histone deacetylases (HDACs), viral proteins, immunophilins, and STAT1.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.