Imaging, protein analysis

Los GV et al (2008) HatoTag a novel protein labeling technology for cell imaging and protein analysis. ACS Chem Biol 3 373-382... 

Los GV, Wood K. (2007) The HaloTag a novel technology for cell imaging and protein analysis Methods Mol Biol 356, 195-208. 

Fig. 24 GST yeast protein analysis, a 60 samples were examined by immunoblot analysis using anti-GST 19 representative samples are shown, b 6566 protein samples representing 5800 unique proteins were spotted in duplicate onto a single nickel-coated microscope slide which was then probed with anti-GST. c Enlarged image of one of the 48 blocks [ 143]...

Liu, Z. and Pawliszyn, J., Coupling of solid-phase microextraction and capillary isoelectric focusing with laser-induced fluorescence whole column imaging detection for protein analysis. Analytical Chemistry, 77,165, 2005. 

As a practical example, when a mouse brain section to which 2,5-dihydroxybenzoic acid (DHB) has been applied as a matrix is subjected directly to MS in positive ion-detection conditions, strong peaks which are mainly derived from phospholipids were observed in mass region of 700 < m/z < 900 [8] on the other hand, signals derived from proteins, meanwhile, are scarcely detected at m/z > 3000 [9].This is because phospholipids ionize much more efficiently than proteins, and they, on the other hand, suppress protein/peptide ionization. Therefore, for detecting/imaging proteins and peptides, removal of such lipids improves the sensitivity for proteins analysis. To this end, tissue sections should be rinsed with organic solvent, to remove lipids from tissue samples [9-11]. 

Nakanishi, T. Shibue, Y. Fukuyama, Y Yoshida, K. Fukuda, H. Shirasaka, Y Tamai, I. Quantitative time-lapse imaging-based analysis of drug-drug interaction mediated by hepatobiliary transporter, multidrug resistance-associated protein 2, in sandwich-cultured rat hepatocytes. Drug Metab. Dispos. 2011, 39,984-991. 

Besides sensitive methods for the analysis of proteins, bioinformatics is one of the key components of proteome research. This includes software to monitor and quantify the separation of complex samples, e.g., to analyze 2DE images. Web-based database search engines are available to compare experimentally measured peptide masses or sequence ions of protein digests with theoretical values of peptides derived from protein sequences. Websites for database searching with mass spectrometric data may be found at http //www.expasy.ch/tools, http //prospector.ucsf. edu/ and http //www.matrixscience.com. 

An assay that produces multiple biological readouts. Most commonly used in relation to the mathematical analysis of an image acquired using an automated microscope whereby analysis algorithms quantify cellular parameters (e.g., number, motility, neurite outgrowth, size, shape) and subcellular events (e.g., receptor internalization, protein translocation, protein expression nuclei shape). 

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