Artifactual immunofluorescent labelling in yeast, demonstrated by affinity purification of antibody

Artifactual immunofluorescent labelling in yeast, demonstrated by affinity purification of antibody.

In the course of making antibodies against various yeast (S. cerevisiae) proteins, we have noted that it is common to observe reactivity of rabbit sera with a number of extraneous bands on Western transfers of yeast proteins. The pattern of reactive bands can change within a period of weeks, even when the rabbit has not been injected with antigen. A simple method of affinity purification, using...

Yeast mitochondrial ADP/ATP carriers are monomeric in detergents as demonstrated by differential affinity purification.

Most mitochondrial carriers carry out equimolar exchange of substrates and they are believed widely to exist as homo-dimers. Here we show by differential tagging that the yeast mitochondrial ADP/ATP carrier AAC2 is a monomer in mild detergents. Carriers with and without six-histidine or hemagglutinin tags were co-expressed in defined molar ratios in yeast mitochondrial membranes. Their specific...

Engineering antibody affinity by yeast surface display.

Yeast surface display (YSD) is a powerful tool for engineering the affinity, specificity, and stability of antibodies, as well as other proteins. Since first described in 1997 by Boder and Wittrup, YSD has been employed successfully in engineering a number of antibodies, as well as T-cell receptors. A recently reported large nonimmune single chain antibody library serves as a good starting poin...

University of Groningen Yeast Mitochondrial ADP/ATP Carriers Are Monomeric in Detergents as Demonstrated by Differential Affinity Purification

Purification of urokinase by monoclonal antibody affinity chromatography.

Matrix-bound monoclonal antibodies against urokinase have been used to purify this enzyme by affinity chromatography. In a single-step procedure, urokinase can be isolated from crude preparations with high yield and high purity, and without loss of enzymatic activity.