Ph.D.: Ben-Gurion University of the Negev, Israel
Post-Doctorate: Ben-Gurion University of the Negev, Israel
Position: Research Associate
ProteOn and FACS Unit-Head
E-mail: azilha@bgu.ac.il

ProteOn and FACS Unit

Background

Surface plasmon resonance (SPR) - ProteOn XPR36

The function of living cells involves the coordination of many thousands of reactions at every level, from the metabolism of small molecules to the signaling processes that lead to cell division or cell death. These different reactions must be integrated to ensure balanced and timely provision of all necessary components and to adjust to changes in environment. To address inter-molecular reactions, the NIBN has recently purchased the ProteOn XPR36 protein interaction array system, an advanced Surface Plasmon Resonance (SPR)-based tool designed for real-time bio-specific interaction analysis. The ProteOn allows for the study of molecular interactions directly and continuously over time without the need for exogenous reagents. Its optical biosensor provides rapid and efficient optimization of experimental conditions by integrating a high-efficiency crisscross microfluidics system with a high-sensitive optical system to generate data within a 6x6 array. This set-up allows for simultaneous measurements of 36 individual molecular interactions as a function of time using specific sensor chips. ProteOn technology can be used to search for the binding partners to a chip-fixed target molecule, be they protein, carbohydrate or oligonucleotide. The system can, moreover, use the same sensor chip for specificity, interaction kinetic, and affinity measurements, amongst others.

Fluorescence-activated cell sorter (FACS) analysis

The FACS relies on microfluidic-based technology for counting, examining and sorting microscopic particles suspended in a stream of fluid. It allows simultaneous multiparametric analysis of the physical and/or chemical characteristics of single cells flowing through an optical detection apparatus using laser light and one or more fluorescent detectors. Using the FACS, several cell parameters can be examined, such as cell volume, the morphological complexity of cells, and cell surface antigens. Cell sorting is also a central application of this technique. FACS can also address apoptosis by quantification and measurement of DNA degradation, mitochondrial membrane potential, permeability changes, and caspase activity. Likewise, DNA analysis during the cell cycle, cell proliferation and apoptosis, is possible. FACS technology, therefore, finds widespread use in a wide range of biological and medical research fields, such as cell biology, tumor immunology, molecular biology, and plant biology. The NIBN is equipped with the FACSvantage SE, proficient in cell sorting and able to simultaneously examine up to 4 different fluorescence markers.

Selected publications

Zilka A., Garlapati S., Dahan E., Yalosky V. and Shapira M. (2001) Developmental regulation of Heat Shock Protein 83 in Leishmania; 3’ processing and mRNA stability control transcript abundance and translation is directed by a determinant in the 3’- untranslated region.

Journal of Biological Chemistry 276(51), 47922-47929.

Zilka A., Landau G., Hershkovitz O., Bloushtain N., Bar-Ilan A., Benchetrit F., Fima E., van Kuppevelt T.H., Gallagher J.T., Elgavish S., Porgador A. (2005) Characterization of the Heparin/Heparan Sulfate Binding Site of the Natural Cytotoxicity Receptor NKp46.

Biochemistry 44(44):14477-14485.

Hershkovitz O, Jivov S, Bloushtain N., Zilka A., Landau G., Bar-Ilan A., Glicklis R., van Kuppevelt T. H., and Porgador A. (2007) Characterization of the recognition of tumor cells bythe natural cytotoxicity receptor, NKp44 Biochemistry 46(25):7426-7436.

Cagnanol E., Hershkovitz O., Zilka A., Bar-Ilan A., Sion-Vardy1 N., Mandelboim O., Benharroch D. and Porgador A. (2008) Expression of ligands to NKp46 in benign and malignant melanocyte. Jurnal of investigative dermatology 128: 972-979.