Elution

Having retained the compound or macromolecule of interest on the immobilized antibody, elution can be accomplished by a variety of methods. If the antibody is covalently bound to the support, relatively strong eluotropic conditions can be used including organic solvents such as ethanol, changes in buffer concentration and/or pH, or the use of chaotropic reagents. For speed and sharp elution profiles of the analyte/prod-uct, a rapid change in eluent composition from conditions promoting retention to those favouring elution can and are used, i.e. a simple step gradient. However, gradient elution can be used if less aggressive conditions need to be employed and the dilution of the target molecule, relative to step gradient elution, can be accommodated. A further method of elution that can be used where antibodies with relatively weak affinities are used is competitive displacement using another molecule that has an affinity for the antibody (this technique has been termed 'weak affinity chromatography'). In addition, as described below in greater detail, peptides (against which the antibodies were raised) that correspond to a particular region of the target proteins can be used to promote desorption.

Furthermore, as noted above, certain proteins binding to the antibody is metal ion dependent. Thus the protein C bound in the absence of calcium ions was eluted from the immunosorbent using a calcium chloride-containing buffer.

Solar Panel Basics

Solar Panel Basics

Global warming is a huge problem which will significantly affect every country in the world. Many people all over the world are trying to do whatever they can to help combat the effects of global warming. One of the ways that people can fight global warming is to reduce their dependence on non-renewable energy sources like oil and petroleum based products.

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