The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Detects a band of approximately 358 kDa (predicted molecular weight: 358 kDa).
Use at an assay dependent dilution.
E3 SUMO-protein ligase which facilitates SUMO1 and SUMO2 conjugation by UBE2I. Involved in transport factor (Ran-GTP, karyopherin)-mediated protein import via the F-G repeat-containing domain which acts as a docking site for substrates. Could also have isomerase or chaperone activity and may bind RNA or DNA. Component of the nuclear export pathway. Specific docking site for the nuclear export factor exportin-1.
Protein modification; protein sumoylation.
Defects in RANBP2 are the cause of susceptibility to encephalopathy acute necrotizing type 1 (ANE1) [MIM:608033]. A rapidly progressive encephalopathy manifesting in susceptibile individuals with seizures and coma. It can occur within days in otherwise healthy children after common viral infections such as influenza and parainfluenza, without evidence of viral infection of the brain or inflammatory cell infiltration. Brain T2-weighted magnetic resonance imaging reveals characteristic symmetric lesions present in the thalami, pons and brainstem.
RanBP2 was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Rabbit polyclonal to RanBP2 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation. Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab64276. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697) . Band: 358kDa: RanBP2.
ab64276 (1:2000) staining RanBP2 in Hela cells pre-extracted in 0.005% digitonin and fixed in 3.7% PFA/30 mM sucrose/PBS for 15 min (room temperature).Image kindly provided by Patrizia Lavia, Univ. "La Sapienza", CNR, Italy.
Western blot - RanBP2 antibody (ab64276)
Anti-RanBP2 antibody (ab64276) at 1/2000 dilution + Hela whole cell lysate
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