pDsRed-Monomer-Hyg-C1 Vector

Schematic for fluorescent fusion protein vectors

Schematic for fluorescent fusion protein vectors. Panel A. N-terminal fluorescent protein (FP) vector. Panel B. C-terminal fluorescent protein vector.

Hygromycin DsRed-Monomer and AcGFP1 expression vectors

Hygromycin DsRed-Monomer and AcGFP1 expression vectors. Vector maps of our DsRed-Monomer and pAcGFP1 Hyg-N1 (Panel A) and Hyg-C1 (Panel B) mammalian expression vectors for fusion applications. Each vector contains a hygromycin selection cassette. These vectors can be used together with our standard N1/C1 vectors containing the neomycin selection cassette to establish double stable cell lines expressing two different Living Colors proteins.

DsRed-Monomer is soluble when expressed in mammalian cells

DsRed-Monomer is soluble when expressed in mammalian cells. HeLa cells were transfected with pDsRed-Monomer-N1 and fixed in 4 paraformaldehyde 24 hr post-transfection. DsRed-Monomer fluorescent protein displays an even, consistent, and homogeneous distribution.

DsRed-Monomer is a monomeric protein

DsRed-Monomer is a monomeric protein. Panel A. Recombinant DsRed-Express and DsRed-Monomer fluorescent proteins (100 μg) were analyzed by FPLC gel filtration chromatography. Overall absorbance (A280) and chromophore excitation (A557) of the eluted material were monitored simultaneously. DsRed-Monomer elutes from the column at a retention time (39 min) corresponding to a molecular weight of 28 kDa. The calculated molecular weight of DsRed-Monomer is 26.8 kDa. DsRed-Express is a tetrameric protein that elutes at an earlier retention time (33 min) corresponding to a molecular weight of 89 kDa. Panel B. 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, DsRed-Monomer fluorescent protein 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 boiled (denatured) counterpart due to its tetrameric structure.

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1

Fluorescence excitation and emission spectra of DsRed-Monomer and AcGFP1.