When white light is passed through cell, it’s hard to see much because of transparency, so you can use DIC or phase contrast.
Phase Contrast - Two waves of light in phase increases intensity of the light and you are able to see better if disrupted, that creates contrast to be visualized by eye.
DIC – disrupts light but put a prism in microscope that bends light to create a bright halo around organelles and vesicles and plasma membrane. Usually used to visualize vesicle transport. Allows you to image live cells.
Epitopes – epitope binding sites. A single antibody recognizes a single epitope on a protein or bacteria. Some epitopes induce an immune response if the region on the protein/bacteria is immunogenic, others will not and those are called antigenic.
A single b cell has a single antibody as well as a single epitope specificity. They are expressed in the membrane.
Taking all proteins in cell, removing them from cell by collecting them all, and running on a polyacrylimide gel, separating by size.
Coomassie stained gels – directly staining the gel to observe separated proteins.
Take a filter to probe with antibody of interest and use to detect whether or not the protein it recognizes is there.
Can use them to bind proteins to see what happens if protein doesn’t function.</li></ul>A secondary antibody is an antibody that binds to primary antibodies or antibody fragments. They are typically labeled with probes that make them useful for detection, purification or cell sorting applications.<br />Secondary antibodies may be polyclonal or monoclonal, and are available with specificity for whole Ig molecules or antibody fragments such as the Fc or Fab regions.<br />Specific secondary antibodies are usually chosen to work in specific laboratory applications. Frequently, any one of several secondary antibodies perform adequately in a particular application. They are selected according to the source of the primary antibody, the class of the primary antibody (e.g., IgG or IgM), and the kind of label which is preferred. Identifying the optimal secondary antibody is normally done through trial and error.<br />