Antibody News

Western blot is a widely used immunoassay technique, used to identify proteins. Many people enter Western blot research without having a clear idea of how the technique relates to antibody usage, so we at Novus Biologicals thought it would be interesting to give a general review of this subject.

Western blotting, also called protein immunoblotting, uses gel electrophoresis to separate proteins of tissue extracts and homogenates. Native and denatured proteins are identified by their isoelectric point, electric charge, molecular weight or molecular structure (often, a combination of these).

Tissue for Western Blot analysis is blended or homogenized to break down cells and release the proteins. Bacterial and viral proteins are also identified using Western blot, and samples are not restricted to cellular tissues...

Recombinant proteins are those derived from recombinant DNA. Recombinant DNA is an artificial form of DNA, made by combining protein sequences which would not occur in their natural state.

Recently, we at Novus Biologicals have been developing recombinant bacterial proteins to reproduce existing products on our antibody database. Recombinant proteins are created from complementary cDNA libraries, which contain only DNA sequences that code for specific proteins. When this cDNA is expressed as a recombinant protein, it can be used in its purified form to perform studies into, for example, enzyme kinetics and NMR. The protein can also be used to create antibodies for further research – something which has been keeping our lab staff busy in recent times!

To take this further, if DNA sequences for certain genes are unavailable or not known, antibodies could be used for screening instead....

A conjugated antibody (also known as a tagged, loaded or labeled antibody) is one that has been attached to a substrate such as an enzyme, toxin or inorganic compound. Modern immunoassay techniques frequently use proteins conjugated with a fluorescent dye. Contemporary antibody sources sometimes supply a wide range of both polyclonal and monoclonal conjugated antibodies. For the benefit of those who are new to this area, we thought it would be a good idea to provide a synopsis of what these proteins actually are, how they work and how they are used. Conjugated antisera are used in many research applications, from Western blotting to in vivo cellular analysis. Researchers are looking for two things – a strong signal and specificity to particular antigens. We at Novus Biologicals pride ourselves on having an extensive catalog of highly specific products, including antibodies tagged with the very latest...

Researchers routinely use GTP binding antibodies for the research of human disease. Thus it is worthwhile to provide a recap of the GTP binding process.

GTPases, a type of heterotrimeric G proteins, are essential to all metabolic functions. They act as molecular switches on the cell membrane, activating in response to various chemical signals. For example hormones, cytokines, physical stresses and cell-to-cell communications all activate downstream pathways. The interactions are complex, each requiring a different GTP protein. These function as molecular switches to activate or deactivate specific cellular responses.

G-proteins work by hydrolyzing GTP (guanosine triphosphate). They switch between active and inactive states, depending on whether they are GTP (guanosine triphosphate) or GDP (guanosine diphosphate) bound. In their active (GTP) state, they interact with effectors to initiate a downstream signal. In their GDP state...