pAcGFP1-Mito Vector

Description

pAcGFP1-Mito encodes a humanized green fluorescent protein GFP derived fromAequorea coerulescens. A mitochondrial targeting sequence from the precursor protein of human cytochrome c oxidase subunit VIII is fused to the N-terminus of the fluorescent protein. The fluorescence from pAcGFP1-Mito expression can be observed within the mitochondrial matrix inside the inner membrane. pAcGFP1-Mito can be used for specific labeling of mitochondria in living and fixed cells. 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 AcGFP1 is ideal for multicolor and fluorescence microscopy applications.AcGFP1 and DsRed2 protein fusions were transiently transfected and visualized by fluorescence microscopy.Panel A.pAcGFP1-Mito mitochondria and pDsRed2-Nuc nucleus in HEK 293 cells.Panel B.pAcGFP1-Golgi Golgi apparatus and pDsRed2-Nuc nucleus in HEK 293 cells. Back Organelles targeted by the subcellular localization vectors. Back Subcellular localization vectors encode fusions of fluorescent proteins with localization signals or subcellular structural proteins, which target the fluorescent protein to a specific organelle or subcellular structure.The vectors are available in a variety of organelle- and cytoskeleton-targeted color variants. Back Use of AcGFP1 for fusions and fluorescence microscopy applications.Panels AandB. Activation of Protein Kinase C alpha was monitored with Living Colors AcGFP1.Panel A. HEK 293 cells were stably transfected with a plasmid encoding AcGFP1 fused to PKC alpha.Panel B. Cells were induced with 1.5 mug/ml PMA for 3 min. The PKC alpha-AcGFP1 fusion moves from the cytosol to the plasma membrane, a result consistent with the known mobilization pattern of PKC alpha.Panel C. HeLa cells were transiently transfected with pAcGFP1-Actin and visualized by fluorescence microscopy.