Purification of Prealbumin from Human and Canine Serum Using a Two-Step Affinity Chromatographic Procedure

Two-step affinity purification of multiubiquitylated proteins from Saccharomyces cerevisiae.

In budding yeast and higher eukaryotic genomes, there are, respectively, 50 and up to 400 or more distinct genes that encode for ubiquitin-ligases, and approximately 15-90 genes that encode for ubiquitin isopeptidases (TM and RJD, Semple et al., 2003). This puts ubiquitylation on par with phosphorylation as the most common reversible posttranslational modifications in eukaryotic cells. A key ch...

Purification of intact old yellow enzyme using an affinity matrix for the sole chromatographic step.

Old Yellow Enzyme has been purified in high yield from crude extracts of brewers bottom yeast using an affinity matrix for the sole chromatographic step. The results of sodium dodecyl sulfate gel electrophoresis indicate that the enzyme is nearly homogeneous and consists of identically sized subunits of molecular weight of about 49,000. However, when the enzyme is isolated without the protectiv...

Requirement of a Blocking Step in Affinity Purification of Polyclonal Antibodies

Purification of egg yolk immunoglobulins. A two-step procedure using hydrophobic interaction chromatography and gel filtration.

A two-step chromatographic procedure was developed for the isolation and purification of hen IgY antibodies from egg yolk. The antibodies were completely separated from vitellin and lipids by hydrophobic interaction chromatography followed by gel filtration. Almost no residual yolk proteins, no immunoglobulin aggregates, and no antibody fragments could be detected in the final extract. Moreover...

Single Step Purification of Novel Thermostable and Chelator Resistant Amylase from Bacillus Licheniformis RM44 by Affinity Chromatography

Bacillus licheniformis RM44 was isolated from hot spring near Karachi and screened forthe production of extracellular amylase Amy RM44. Amy RM44 was purified to homogeneityon a single step by affinity chromatography using insoluble corn starch. The molecular weightof Amy RM44 was estimated to be 66 kDa by SDS–PAGE and zymographic analysis. Nine foldpurification was achieved with the specific ac...