pIRES2-AcGFP1-Nuc Vector
pIRES2-AcGFP1-Nuc Vector
Brand: Takara Bio.
In stock
SKU
pIRES2-AcGFP1-Nuc Vector
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.
AcGFP1 is a monomeric protein
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 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) 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.
Map of the fluorescent pIRES2 bicistronic expression vectors
Map of the fluorescent pIRES2 bicistronic expression vectors.
Expression of two proteins from a single mRNA transcript
Expression of two proteins from a single mRNA transcript. A fluorescent protein is translated from an internal ribosome entry site (IRES).
Write Your Own Review
AcGFP1 is a monomeric protein
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 4°C and not boiled prior to loading on a gel. Boiled and unboiled recombinant proteins (7.5 μg) 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.
Map of the fluorescent pIRES2 bicistronic expression vectors
Map of the fluorescent pIRES2 bicistronic expression vectors.
Expression of two proteins from a single mRNA transcript
Expression of two proteins from a single mRNA transcript. A fluorescent protein is translated from an internal ribosome entry site (IRES).
