pIRES2-AcGFP1-Nuc Vector

Description

pIRES2-AcGFP1-Nuc encodes AcGFP1 with three identical copies of the nuclear localization signal NLS of the simian virus 40 large T antigen fused in tandem to its C-terminus. The gene encoding the AcGFP1-Nuc fusion protein is located downstream of the IRES2 sequence. This permits both the gene of interest cloned into the MCS upstream of the IRES2 sequence and the AcGFP1-Nuc gene to be translated from a single bicistronic mRNA. AcGFP-1-Nuc localizes to the nucleus of a transfected cell, offering an advantage over random localization in the cytosol. It is most useful for those who are interested in monitoring translocation events in and out of the nucleus. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back AcGFP1 is a monomeric protein.Panel A. Recombinant AcGFP1 protein was analyzed by FPLC gel filtration chromatography. Overall protein absorbance A280 and chromophore excitation A477 of the eluted material were monitored simultaneously. AcGFP1 elutes from the column at a retention time corresponding to a molecular weight of 24 kDa. The calculated molecular weight of AcGFP1 is 26.9 kDa.Panel B. Recombinant AcGFP1 protein was analyzed by sucrose density ultracentrifugation using a continuous gradient.Panel C. Pseudonative gel analysis of proteins. The oligomeric structure of proteins is preserved during SDS-PAGE analysis if samples are kept at 4C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins 7.5 mug were separated by SDS-PAGE electrophoresis 12 % acrylamide . In both the boiled denatured and unboiled nondenatured samples, AcGFP1 runs as a uniform band of 30 kDa due to its monomeric structure. The unboiled nondenatured DsRed-Express runs at a much higher molecular weight than its denatured boiled counterpart due to its oligomeric structure. Back Back Back