Likewise, serum proteins like glycoproteins and albumin are also analyzed using this technique. Blood analysis: The cellulose acetate electrophoresis is applicable to determine hemoglobin abnormalities, especially in people with undiagnosed sickle cell anemia.The applications of cellulose acetate electrophoresis are: So, it is sometimes referred to as hemoglobin electrophoresis. Lane 1: homozygous sickle cell lane 2: β-thal trait and Hb X lane 3: proband lane 4: sickle cell trait control lane 5: normal adult control. B) Isoelectric focusing on polyacrylamide gel. Lane 1: normal adult control lane 2: proband lane 3: sickle cell trait control lane 4: homozygous sickle cell. A) Cellulose acetate electrophoresis at alkaline pH. The most common application of cellulose acetate electrophoresis is in the diagnostic area for determining abnormalities in human hemoglobin.Įlectrophoretic study.

Application of Cellulose Acetate Electrophoresis

Spectrophotometer/zymogram paper/ materials for blotting techniques are required for detecting and quantifying the separated components.

Filter paper wicks: 3mm Whatman filter paper wick with a length similar to the width of an electrophoretic tank.

Power supply: A power supply that can deliver constant current is required.

Electrophoresis tank: An horizontal electrophoresis tank with an adjustable bridge is used.

Electrophoresis buffer: Tris-EDTA buffer (TEB) of 8.4 pH is usually used for analyzing blood.

Here one side of the strip acts as the cathode and another as the anode. The cellulose paper is acetylated by treating it with acetic anhydride.

Cellulose acetate strip/membrane: Cellulose acetate is a thermoplastic resin.

Horizontal cellulose acetate electrophoresis unit Materials Required for Cellulose Acetate Electrophoresis Other methods of semi-quantifying the separated band are treating it with an appropriate blotting technique or using a spectrophotometer. The strips are peeled away from the filter paper the zymogram paper is treated as per the required chemicals for suspected proteins.

The strip is placed in filter paper, soaked in buffer and substrate, and incubated for one hour. The separated band is detected using the zymogram method after applying current for the desired time. (1) The positively charged substance moves to the cathode, and the negatively charged substance moves to the anode. The components are separated in the form of a band based on their charge. The electric volt of 6-8 V is provided in the strip for about an hour. The end of the strip is in contact with the buffer with the help of a filter paper wick. Then the sample is loaded in the strip (approximately ⅓ area of the strip). The cellulose acetate strip is first wetted in the electrophoretic buffer. (1)ĭiagram of cellulose acetate electrophoresis The separation of proteins and peptides in the cellulose acetate occurs based on the isoelectric point of the components, where the overall charge of proteins and peptides are zero. This property helps to analyze constituents of similar size easily using this technique. After all, separation is not based on the size of the component. It is a suitable support matrix because it has larger pores than the agarose or polyacrylamide gels. The cellulose acetate is obtained from the acetylation of cellulose paper (filter paper). The principle of cellulose acetate electrophoresis is similar to the general principle of electrophoresis, where a support matrix (cellulose acetate strip) is used to separate sample components in a buffer system under electric volt. Limitations Principle of Cellulose Acetate Electrophoresis.Application of Cellulose Acetate Electrophoresis.Materials Required for Cellulose Acetate Electrophoresis.Principle of Cellulose Acetate Electrophoresis.

Electrophoresis cathode charge