This chapter discusses the equilibrium and kinetic inhibition assays based upon fluorescence polarization. Assays, utilizing the high specificities of antibodies or other receptors make it possible to detect and quantify many substances present in only minute traces in complex biological materials. The polarization of fluorescence operates differently than the other methods of detection. In that, it gives a direct measure of the bound/free ratio instead of simply being a measure of amount of the label present. Fluorescence polarization experiments can be made, by observing, either the steady state, resulting from constant illumination of the sample or the transient state that follows a very short pulse of light. An analysis is made of the rates of the reactions that occur after preformed complexes are rapidly diluted so that the complexes dissociate to some extent and approach a new equilibrium state. The dilution jump assay affords a means of distinguishing between the specific and nonspecific interactions provided that the specific and nonspecific complexes dissociate with different rates. The assays, utilizing chaotropes to distinguish between the specific and nonspecific binding, are also discussed in the chapter. Series: Methods in Enzymology (Book 74) Hardcover: 736 pages Publisher: Academic Press; Language: English ISBN-10: 0121819744 ISBN-13: 978-0121819743 Product Dimensions: 6 x 9 inches Link Download http://nitroflare.com/view/A94C8E030398803https://drive.google.com/drive/folders/1yLBzZ1rSQoNjmWeJTZ3WGQHg04L1
