The movement of scattered particles within a fluid under the effect of a relatively uniform electric field is known as electrophoresis. Cataphoresis is called the electrophoresis of cations (positively charged particles), whereas anaphoresis is the electrophoresis of anions (negatively charged particles).

Gel electrophoresis segregates DNA fragments (and other macromolecules, such as proteins and RNA) based on their charge and size. A gel containing the target molecules is subjected to electrophoresis by passing an electrical current. The molecules will move through the gel at different speeds depending on their size and charge, enabling them to be isolated from one another.

1D-Gel Electrophoresis
2D-Gel Electrophoresis
Difference between 1D-Gel Electrophoresis and 2D-Gel Electrophoresis
Frequently Asked Questions (FAQs)

1D-Gel Electrophoresis

One-dimensional gel electrophoresis, also referred to as 1D-Gel electrophoresis, is a method for separating proteins depending on their molecular weight. Polyacrylamide gel electrophoresis is mostly used for protein separation.

Proteins can be separated using 1D-Gel electrophoresis by molecular weight across a range of approximately 10 kDa to 300 kDa (kilodaltons). This procedure involves weighing and dissolving materials in sodium dodecyl sulphate (SDS). SDS is a negatively charged detergent with hydrophilic (associated with water) and hydrophobic (not associated with water) parts.

Proteins are segregated using 1D-Gel electrophoresis as per their molecular weight. Thus, the lesser-weight molecules move through the gel more quickly than the proteins with larger molecular weights. As a result, high-weight proteins remain closer to wells.

2D-Gel Electrophoresis

Proteins are separated using two-dimensional gel electrophoresis, often known as 2D-Gel electrophoresis, based on the protein’s isoelectric point and molecular weight. Protein separation resolution is improved by using this technique. We can determine its isoelectric point based on the pH level at which a protein becomes neutral.

In 2D-Gel electrophoresis, the protein moves over a set pH gradient at first. Proteins are then divided using polyacrylamide gel electrophoresis in either a vertical or horizontal pattern. As a result, in the second dimension, proteins segregate based on their molecular weight.

Additionally, the resolution of protein separation is improved by using this gel electrophoresis method. Thus, proteins that have been isolated are purer. The cost of the method is significantly higher than that of one-dimensional gel electrophoresis.

Difference between 1D-Gel Electrophoresis and 2D-Gel Electrophoresis

1D-Gel Electrophoresis	2D-Gel Electrophoresis
It is a process of protein separation based on the molecular weight utilising polyacrylamide gel electrophoresis.	It segregates proteins based on the protein’s molecular weight and iso-electric point.
Protein separation is based on molecular weight only.	Protein separation is based on molecular weight and iso-electric point of the protein.
Low-resolution	High-resolution
Not so costly	Very costly

Protein characterisation depends primarily on protein separation by gel electrophoresis. Compared to DNA segregation by Agarose gel electrophoresis, proteins have diversified characteristics, making their separation more challenging.

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Frequently Asked Questions – FAQs

What is 2D-Gel electrophoresis used for?

2-DE or 2-D electrophoresis, known as two-dimensional gel electrophoresis, is a type of gel electrophoresis often used to examine proteins. Mixtures of proteins are segregated by two attributes in two dimensions on 2D-Gels. In 1975, O’Farrell and Klose independently developed 2-DE for the first time.

What is the difference between gel electrophoresis and SDS PAGE?

The major distinction between gel electrophoresis and SDS PAGE is that while SDS PAGE is a form of gel electrophoresis that is primarily used to separate proteins, gel electrophoresis is a procedure for separating DNA, RNA and proteins. SDS PAGE usually provides a higher resolution than conventional gel electrophoresis.

How does polyacrylamide gel electrophoresis work?

An effective method for examining RNA samples is polyacrylamide gel electrophoresis. Smaller molecules can enter and pass through the pores of polyacrylamide gel, making it easier to study smaller molecules, whereas larger molecules are stuck at the pore openings.

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