Cell-Free Protein Expression: Characterization of Plant Proteins

Cell free protein expression can be utilized for the analysis of: protein/protein interactions, protein nucleic acid interactions, analysis of post translational modifications and many other applications. The majority of these references are based on the characterization of mammalian proteins.
However there are several references using TNT-based systems (either rabbit reticulocyte lysate or wheat germ based) for the analysis of proteins from plants, examples include: Continue reading “Cell-Free Protein Expression: Characterization of Plant Proteins”

Cell-Free Expression Applications: Characterization of Viral-Mediated Diseases

INFECTION IS BY TRANSCRIPTION OF THE VIRAL DNA INTO THE GENETIC MATERIAL OF THE HOST CELL, SO ANTIBODY TEST ING CAN DIAGNOSE AIDS. MORE SIGNIFICANTLY, IF THE PERCENTAGE OF LYMPHOCYTES THAT ARE CD4+ IS ABOVE 500 PER CUBIC MM THERE IS BETTER PROGNOSIS FOR TREATMENT. HERE THE VIRUS IS BUDDING FROM THE T CELL AS PART OF ITS PROLIFERATION AFTER INFECTING THE T CELL. AIDS VIRUS DIGITIZED TEM COLOR ENHANCED

The characterization of viral mediated diseases is critical to promote the overall welfare of humans or animals. Initial research focused on the interpretation the genomic content (i.e., DNA or RNA based) of the selected virus. The next step is to better understand the proteins that are encoded by this content and their interaction with the host proteome.
The following citations illustrate the use of cell-free protein expression to facilitate this research. Continue reading “Cell-Free Expression Applications: Characterization of Viral-Mediated Diseases”

Enhanced In-Solution Tryptic Digestions: Immobilized Trypsin

In many proteome studies, sample analysis is performed directly from the liquid phase. This method avoids many of the time consuming steps associated with in-gel digestion. As with in-gel digestions, trypsin is the enzyme of choice. Trypsin cleaves at C-terminal of lysine (K) and arginine (R) amino acids resulting in a N-terminal amine group that can accept a proton and a basic side chain residue of the K/R that will also take up a proton. These charged particles can be easily ionized when analyzed by mass spectrometry.

Immobilized Trypsin provides a fast and convenient method for digesting a range of concentrations of purified protein, or complex protein mixtures. Continue reading “Enhanced In-Solution Tryptic Digestions: Immobilized Trypsin”

Create Custom Microarrays for Your Research Needs

In the post-genome sequencing era, researchers are increasingly turning their attention to the proteins encoded within the genome. How are their synthesis, degradation and conformation regulated? Do they interact with other proteins or nucleic acids or lipids? Can these interactions be perturbed? How do changes in the coding sequence of the gene affect the proteins and their function? Like DNA microarrays, protein arrays fulfill a need for miniaturization and throughput, but immobilizing proteins in a way that preserves function and conformation is not a simple problem to solve. Continue reading “Create Custom Microarrays for Your Research Needs”