Cleavable Affinity Purification (Cl-AP): A One-step Procedure to Affinity Purify Protein Complexes

Affinity purification of protein complexes.

INTRODUCTION Knowledge of the composition of protein complexes provides key insights into their functions. Immunoaffinity purification provides an effective means for isolating protein complexes and elucidating their composition. Immunoisolation is achieved with antibodies directed either specifically against the proteins of interest or against tags that are coupled to the proteins of interest....

Lectin-based affinity tag for one-step protein purification.

The production of pure protein is indispensable for many applications in life sciences, however protein purification protocols are difficult to establish, and the experimental procedures are usually tedious and time-consuming. Therefore, a number of tags were developed to which proteins of interest can be fused and subsequently purified by affinity chromatography. We report here on a novel lect...

One-step affinity purification protocol for human telomerase

Human telomerase is a ribonucleoprotein (RNP) enzyme, comprising protein components and an RNA template that catalyses telomere elongation through the addition of TTAGGG repeats. Telomerase function has been implicated in aging and cancer cell immortalization. We report a rapid and efficient one-step purification protocol to obtain highly active telomerase from human cells. The purification is ...

Requirement of a Blocking Step in Affinity Purification of Polyclonal Antibodies

Affinity purification of FLAG - tagged protein complexes : A cautionary tale

Through two simple changes in the composition of the fixative for freeze-substitution of cryoimmobilized specimens, we are able to ameliorate the reproducible visibility of the plasma membrane in D. melanogaster embryos and larvae. The ultrastructural analysis of membrane-defective phenotypes of embryos fixed by our modified method should allow us a refined view on the plasma membrane and give ...