The Protein Man's Blog | A Discussion of Protein Research

Rough handling and proteins do not necessarily go well together. As proteins are extremely sensitive and can easily be degraded during the process of concentration, purification, and storage, they need to be handled with utmost care to reduce, if not eliminate, the risk of unwanted aggregation and/or degradation.

A blocking buffer is a solution of irrelevant protein, mixture of proteins, or other compound that passively adsorbs to all remaining binding surfaces of the plate. The blocking buffer is effective if it improves the sensitivity of an assay by reducing background interference and improving the signal-to-noise ratio. The ideal blocking buffer will bind to all potential sites of nonspecific interaction, eliminating background altogether, without altering or obscuring the epitope for antibody binding.

Introduction

While agarose beads have been traditionally used for immunoprecipitation (IP) and other micro-scale protein and antibody purification procedures, magnetic beads have since taken lead and are now considered tool for protein immunoprecipitation.