By Senta Reichelt (eds.)
The target of this version is to introduce the newbie to the fundamentals of affinity chromatography and supply useful wisdom for the advance of affinity separation protocols. Affinity Chromatography: equipment and Protocols, 3rd Edition courses readers via new cutting-edge protocols, molecular modelling, and the learn of ligand-target interactions. Written within the profitable Methods in Molecular Biology sequence layout, chapters contain introductions to their respective subject matters, lists of the mandatory fabrics and reagents, step by step, simply reproducible protocols, and notes on troubleshooting and fending off recognized pitfalls.
Authoritative and simply accessible, Affinity Chromatography: equipment and Protocols, 3rd Edition is designed as an invaluable source for these attracted to the speedy and quantitative isolation of biomolecules with excessive purity.
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Additional info for Affinity Chromatography: Methods and Protocols
Mix the agarose with the elution buffer by inverting the capped column. Incubate the agarose in elution buffer for at least 10 min at 4 C. 9. Remove the column’s plug and cap. Allow the PAT-containing elution to drain into a collection tube. Store the elution at 4 C. Save a small sample for SDS-PAGE analysis. 10. Analyze collected fractions from above steps by SDS-PAGE (see Note 9). Load 10 μl of each sample on SDS-PAGE gel (Fig. 1). Fig. 1 SDS-PAGE of PAT purified from E. coli lysate using Reactive brown 10-agarose.
Anal Chem 73:198A–205A 96. Moser AC, Hage DS (2006) Chromatographic immunoassays. In: Hage DS (ed) Handbook of affinity chromatography, 2nd edn. Taylor and Francis, Boca Raton, Chapter 29 97. Allenmark S (1991) Chromatographic enantioseparation: methods and applications, 2nd edn. Ellis Horwood, New York 98. Patel S, Wainer IW, Lough WJ (2006) Affinity-based chiral stationary phases. In: Hage DS (ed) Handbook of affinity chromatography, 2nd edn. Taylor and Francis, Boca Raton, Chapter 21 99. Wainer IW (ed) (1993) Drug stereochemistry: analytical methods and pharmacology, 2nd edn.
Yoo MJ, Hage DS (2011) Use of peak decay analysis and affinity microcolumns containing silica monoliths for rapid determination of drug-protein dissociation rates. J Chromatogr A 1218:2072–2078 129. Jozwiak K, Haginaka J, Moaddel R, Wainer IW (2002) Displacement and nonlinear chromatographic techniques in the investigation of interaction of noncompetitive inhibitors with an immobilized α3β4 nicotinic acetylcholine receptor liquid chromatographic stationary phase. Anal Chem 74:4618–4624 130. Moaddel R, Wainer I (2007) Conformational mobility of immobilized proteins.
Affinity Chromatography: Methods and Protocols by Senta Reichelt (eds.)