Differentiation of serum proteins by electrophoresis is important for the diagnosis and prognosis of numerous diseases. Use of serum protein electrophoresis is a common tool in clinical diagnosis to express the various infectious states. Staining of serum proteins in electrophoresis is really an uphill task to define accurate abnormal proteins. Many stains have high sensitivity but it cannot be affordable in common laboratories. Hence, the objective of this comparative study is to establish a standard staining protocol with an appropriate stain with affordable cost and high sensitivity. Amido black is one among the best stains used in older protocols. Due to the influx of new sensitive stains like Ponceau S the use of Amido black has become very less. In our experience, we found Amido black as a better stain in comparison to Ponceau S after many replicates of the experiment.