Gel Electrophoresis is one of the zone electrophoresis, and the supporting medium of it is gel. The gel is a cross-linked polymer, acting as a molecular sieve. Its composition and porosity are chosen based on specific requirements. The types of gel typically used are starch, agarose, and polyacrylamide gels.
Starch: Partially hydrolyzed potato starch is a non-toxic medium for the separation of non-denatured proteins.
Agarose: Agarose is a purified uncharged polysaccharide extracted from seaweeds. By manipulating the concentration of agarose in the gel, the pore size can be adjusted. Agarose gel electrophoresis is suitable for the separation of proteins larger than 200 kDa, and DNA fragments of 0.2-20kb.
Polyacrylamide: Polyacrylamide gel is tougher than agarose gel and is relatively inert, thermostable, strong, and transparent, with a uniform pore size. Polyacrylamide gel electrophoresis (PAGE) is suitable for the separation of proteins from 5 to 2,000 kDa.
Sodium dodecyl sulfate-polyacrylamide: Sodium dodecyl sulfate (SDS) is added to PAGE. SDS-PAGE eliminates the influence of structure and charge in electrophoresis, and proteins are separated based on the differences in their molecular weight. It is suitable for the separation of proteins from 5 to 250 kDa.
Fig.1 Illustration of gel electrophoresis
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Preparation
Prepare electrophoresis buffer with a certain pH. Prepare a gel solution of appropriate concentration, taking care not to produce bubbles, and use a cast to solidify the solution into a gel. -
Sample Application
Put the prepared gel with a cast into the electrophoresis tank, and add an appropriate amount of buffer. Pipette the sample into the sample wells. -
Electrophoresis
Adjust the appropriate distance between the electrodes. Switch on the electrophoresis apparatus and set the voltage and current. Perform electrophoresis for a period of time. -
Determination
After electrophoresis, stain the gel for visible results. Observe, record and analyze the position of separated bands. Or, utilize an imaging system for analysis. -
Downstream processing
After separation, an additional method is often applied to the separated bands for further processing. For example, cut the band out of the gel as a slice to dissolve and purify it.
Notes: Part of the solution involved in this experiment is toxic. Please handle it carefully with gloves.
- Sharp zones
- Better resolution
- Good recovery of proteins and reasonable yield
- Less adsorption of proteins
- Less time consumption
- Easy preparation
- Stability of gel over a wide range of pH and temperature
- Downstream processing
- Separation of proteins
- Separation of DNA fragments for extraction or purification
- Estimation of the DNA concentration and the size of DNA molecules
- Estimation of molecular weight of nucleic acids and proteins
- Analysis of PCR products
- Identification of disulfide bonds between proteins
- Checking the purity of proteins
- Separation or characterization of metals or metal oxide nanoparticles
Electrophoresis Systems
Electrophoresis Accessories
pH Meters
Gel Drying Equipment
Pipettes
Staining Appliances
Transilluminators
Digital Gel Imaging Systems
Chemiluminescence Imaging Systems
Fluorescent Imaging Systems
STEMart provides you with a variety of electrophoresis equipment or consumables to meet your various R&D and application needs. If you have any questions or requirements for gel electrophoresis, please feel free to contact us.