Book ChapterDOI
Affinity chromatography : general methods
Reads0
Chats0
TLDR
Affinity chromatography is one of the most powerful procedures that can be applied to protein purification and the solvent system chosen for the entire affinity chromatography separation is also a critical factor to a good separation.Abstract:
Publisher Summary Affinity chromatography is one of the most powerful procedures that can be applied to protein purification. The solvent system chosen for the entire affinity chromatography separation is also a critical factor to a good separation. The solvent should not degrade the sample. After choosing the affinity gel type, the resin (gel) is prepared for use. The manufacturer usually supplies the instructions needed to prepare the gel correctly. If the gel is supplied in a preswollen state, reconstituting the gel is unnecessary to obtain the full swollen volume. A wash is all that is usually required. The swollen gel is typically supplied in glycerin or similar material, which is used to help in the gel preparation and to stabilize the ligand or activated coupling complex. If the gel needs to be swollen to regain full working volume, then a swelling buffer is used prior to washing. This buffer is often a low concentration phosphate buffer (0.1 M) at or near neutral pH. Swelling times vary between 15 min and 1 hr. After swelling, the gel is washed in the buffer solution used for swelling, distilled water, or starting buffer.read more
Citations
More filters
Journal ArticleDOI
Liquid chromatography: theory and methodology.
TL;DR: Books, Reviews, and Symposia Proceedings 515R Theory and Optimization 516R Theory 517R Optimization 540R Data Analysis 520R Stationary Phase Classification 520R Retention/Solute-Stationary Phase Interactions 521R Modeling/Peak Characterization/Smoothing 521 r Calibration and Curve Fitting
Book ChapterDOI
Affinity chromatography: general methods.
TL;DR: Factors which are important to consider when selecting the ligand, proper attachment chemistry, and the matrix are discussed.
Journal ArticleDOI
Purification of the Xenopus laevis double-stranded RNA adenosine deaminase.
Ronald Hough,Brenda L. Bass +1 more
TL;DR: A double-stranded RNA adenosine deaminase that catalyzes the conversion of adenosines to inosines in duplex RNA substrates was purified to near homogeneity from Xenopus laevis eggs, suggesting that the enzyme may dimerize or associate with other cellular components.
Journal ArticleDOI
Separation and Analysis of Peptides and Proteins
Cynthia K. Larive,Susan M. Lunte,Min Zhong,Melissa D. Perkins,George S. Wilson,Giridharan Gokulrangan,Todd D. Williams,Farhana Afroz,Christian Schöneich,Tiffany S. Derrick,and C. Russell Middaugh,Susan Bogdanowich-Knipp‡ +11 more
Patent
Cytotoxic Compounds and Conjugates
TL;DR: In this paper, the authors present cytoxic compounds useful as drugs or prodrugs and to drug-cleavable substrate conjugates where the drug and substrate are optionally linked through a self-immolative linker.
References
More filters
Journal ArticleDOI
Single-step purification of avidin from egg white by affinity chromatography on biocytin-Sepharose columns
Pedro Cuatrecasas,Meir Wilchek +1 more
Journal ArticleDOI
Some parameters relevant to affinity chromatography on immobilized nucleotides.
TL;DR: The suitability of cellulose and Sepharose as supports for affinity chromatography of two groups of cofactor-linked enzymes, dehydrogenases and kinases, was examined andSepharose was found to be superior.