Traditional immunoassays such as the enzyme-linked immunosorbent assay (ELISA) are able to measure the presence or absence of only one analyte per reaction. Multiplex immunoassays measure dozens of different analytes in a single reaction. This is particularly beneficial for precious samples, and when only a small volume is collected for analysis. The multiplex immunoassay also saves working time since multiple assays can be completed simultaneously.
Immunoprecipitation (IP) is a method used to purify target proteins from whole tissue or from cell culture. There are different types of IP: Single protein, Co-immunoprecipitation (co-ip), Chromatin immunoprecipitation(CHIP), RNA immunoprecipitation (RIP), and tagged protein immunoprecipitation.
Immunoaffinity chromatography is a method for separating target antibodies or antigens from a heterogenous solution. It is column-based, which means that the solution is flowed through a column and eluted at the other end. The column is pre-functionalized with the capture antibody or antigen. The target protein is adsorbed onto the resin-bound capture protein and is retained in the column while the remaining solution is eluted. The fraction containing the target protein is later eluted and purified.
Size exclusion chromatography is used to separate molecules by size, molecular weight, and hydrodynamic volume. The technique can be used with proteins, polymers, and other macromolecules. It can also be used for buffer exchange or desalting a sample. The principle behind size exclusion chromatography is simple, but the technique only works when the correct resin-bound column is matched to the experimental goal.
Enzymes are used in the food, agricultural, cosmetic, and pharmaceutical industries to control and speed up reactions in order to quickly and accurately obtain a valuable final product. Enzymes are crucial to making cheese, brewing beer, baking bread, extracting fruit juice, tanning leather, and much more. Enzymes are also increasingly being used in the production of biofuels and biopolymers. The enzymes can be harvested from microbial sources or can be made synthetically. Yeast and E. coli are commonly engineered to overexpress an enzyme of interest. This type of enzyme engineering is a powerful way to obtain large amounts of enzyme for biocatalysis in order to replace traditional chemical processes.
Proteins are essential components of cells, tissue, and organisms. These macromolecules are made of long strings of amino acids arranged specifically into three dimensional configurations. The side chains of these 22 amino acids create pockets of potential for chemical interactions as the polypeptides fold into their tertiary structures and interact with each other. Proteins initiate and mediate the thousands of biochemical pathways that govern an organism’s function. The careful study of proteins can reveal information about the function of our bodies, the pathways of disease, and the expression of the genetic code.
Nano gold is another name for gold nanoparticles. These nanoparticles are a fraction of the size of human hair and are less than 100 nm in diameter. Nano gold particles are so small that it they are generally found as a colloidal solution, which means that the gold nanoparticles are suspended in a liquid buffer. Therefore, nano gold, or gold nanoparticles are also called colloidal gold. Also, nano gold is generally found in a colloidal solution because gold nanoparticles are created by citrate synthesis. This process involves mixing solutions together to result in the precipitation of gold nanoparticles into solution.
The force of gravity will cause sedimentation of particles from a heterogeneous mixture;larger and denser particles sedimentfaster than the smaller and less dense particles. This phenomenon is useful for separating heterogeneous solutions into independent components, and for the isolation and enrichment of target molecules, cells, and cell organelles. Centrifugation accelerates the separation process by introducing centripetal forces many times greater than gravity. The precipitated particles form a pellet at the bottom of the tube during centrifugation. The rate of sedimentation is dependent on the size and density of the particles, so centrifugation can be used to isolate target particles simply by controlling centrifugal force or the rate of centrifugation. The rate of centrifugation is reported as angular velocity by revolutions per minute (rpm) or as acceleration(g). RPM is dependent on the radius of the rotor in the centrifuge.