Molecular Biology

Biochemical and molecular methods include analysis of RNA, DNA, proteins, selected enzymes by real-time PCR, Western blotting, oxidative analysis, RIA methods, spectroscopic methods and others.

Enzyme assays: to identify the presence and activity of a specific enzyme in the tissues, different enzyme assay methods including spectrophotochemistry, colorimetry, immunoassay, or radiation biology are used in our experimental studies. For more details see: Enzymatic Assay

Real-time PCR (RT-PCR), one of the most widely used methods of gene quantitation, is the technique  of collecting data throughout the PCR process as it occurs, thus combining amplification and detection into a single step. This is achieved using a variety of different fluorescent chemistries that correlate PCR product concentration to fluorescence intensity. Reactions are characterized by the point in time (or PCR cycle) where the target amplification is first detected. This value is usually referred to as cycle threshold (Ct), the time at which fluorescence intensity is greater than background fluorescence. Consequently, the greater the quantity of target DNA in the starting material, the faster a significant increase in fluorescent signal will appear, yielding a lower Ct. For more details see: Real-time PCR for mRNA quantitation

We usually use RT-PCR for detecting gene production involved in NO/ROS balance like NO synthase isoenzymes (eNOS, phosphorylated eNOS, nNOS, iNOS) or antioxidant enzymes like SOD isoenzymes and others.

Western Blot (WB) is a common method to detect and analyze proteins. It is built on a technique that involves transferring, also known as blotting, proteins separated by electrophoresis from the gel to a membrane where they can be visualized specifically. For more details see: Western Blot

We usually use WB for detecting proteins involved in NO/ROS balance like NO synthase isoenzymes (eNOS, phosphorylated eNOS, nNOS, iNOS), antioxidant enzymes like SOD isoenzymes, inflammatory markers e.g. NF-kappaB and others.