Vertical Electrophoresis Tank
Specifications Gel Tray (W*L) 100*100mm Gel Trays(W*L): 82*88mm Comb Thickness 0.75 – 1.0 – 1.5mm Comb Throughput 11 – 15 samples Amount of Gel: 1~2 piece […]
|Gel Tray (W*L)||100*100mm|
|Comb Thickness||0.75 - 1.0 - 1.5mm|
|Comb Throughput||11 - 15 samples|
|Amount of Gel:||1~2 piece|
|NO. of Samples||11~30|
|External Size (L*W*H)||150*120*115mm|
1 x Body tank with the electrode.
1 x Gel casting base.
4 x Notched glass plates.
2 x 0.75mm bonded glass spacer.
2 x 1.0mm bonded glass spacer.
2 x 1.5mm bonded glass spacer.
2 x 0.75 mm thickness11 wells comb.
2 x 1.0mm thickness11 wells comb.
2 x1.5mm thickness 11 wells comb.
2 x 0.75mm thickness 15 wells comb.
2 x 1.0mm thickness 15wells comb.
2 x1.5mm thickness 15wells comb.
2 x Clamp version.
1 x Dummy plate
Introduction and Principle
Vertical electrophoresis tanks are primarily used for nucleic acid and protein sequencing via vertical gel electrophoresis. In a vertical electrophoresis system, the gel matrix is cast vertically in a discontinuous buffer system, and two chambers, i.e., a top chamber and a bottom chamber, are employed.
The vertical electrophoresis tanks employ a discontinuous buffer system comprising a top and a bottom chamber. The top chamber contains a cathode, and the bottom contains an anode. Electrodes present in both chambers generate the required electric field. A thin gel of less than 2mm is poured between two glass-mounted plates so touch that the upper part of the gel is submerged in the upper buffer chamber while the bottom of the gel is immersed in the lower buffer chamber. As soon as the electric field is applied, a small amount of buffer migrates through the gel from the upper chamber towards the lower chamber. As the buffer moves via the gel only, the experimenter has full control over the voltage gradients throughout the experiment. A vertical gel electrophoresis tank allows the researchers to achieve a more effective separation with greater resolution.
Unlike horizontal electrophoresis, vertical systems mostly use polyacrylamide gels with a smaller pore size (about 10-200nm). Acrylamide gel cannot be used for horizontal gel electrophoresis. The gel in the horizontal system must interact with environmental oxygen, and exposure of acrylamide to oxygen can inhibit acrylamide polymerization, thereby interfering with gel formation. In vertical electrophoresis, the buffer only migrates through the gel, and individual compartments do not get exposed to oxygen.
Moreover, acrylamide concentration controls the gel’s pore size and the degree of crosslinking. Pore size decreases as the total acrylamide concentration increases, and the amount of cross-linker is determined relative to the total acrylamide amount. On the other hand, pore size is parabolically related to crosslinking, i.e., at high and low cross-linking, pores are larger in size, whereas they are minimum in size at 4% crosslinking.
Apparatus and Equipment
The vertical electrophoresis tank is made of acrylic and has 140x100x150mm dimensions. It comprises an upper and a lower buffer chamber. The apparatus is equipped with two gel-cast frames and glass plates. The glass plates are accurately aligned to prevent gel leakage. There are two combs for creating wells. The built-in ridges in the combs isolate gel from the air and avoid polymerization reaction in the gel. Gel cast frame, gel cast stand, and electrophoresis core with high-quality and unique design are the features of the vertical electrophoresis tank offered by Conduct Science.
Protein Fraction Identification from Cow’s Milk Casein Complex
Iukalo (2015) developed a vertical electrophoresis system to identify cow milk’s casein complex proteins. The researchers isolated individual casein protein fractions using preparative electrophoresis. They used a vertical electrophoresis tank to determine total casein’s fractional composition and homogeneity of its fractions. The polyacrylamide gel electrophoresis apparatus they used was Studier type, and it allowed change in the chambers’ geometric dimensions and the number of samples for simultaneous analysis. The electrophoresis system was optimized by reducing the dimensions of the electrophoresis chamber. The changeable dimensions of the electrophoretic chamber significantly reduced the time for identifying casein fractions. The scientists concluded that the method they developed could be useful for the robust analysis of milk and dairy products.
Use in Western Blotting for the Detection of TLR4 Expression in Spinal Cord Tissue
Han et al. (2015) studied the effect of bone marrow mesenchymal stem cell (BMSC) on TLR4 (toll-like receptor 4) levels in four-week-old male SD rats. BMSCs were isolated and identified by immunocytochemistry. Then the scientists prepared animal models for BMSC transplantation, prepared spinal cord injury sites, and detected the presence of TNF-α and IL-1β using the RT-PCR method. A vertical electrophoresis tank was used for western blotting to detect TLR4 expression in the spinal cord tissue. In addition, about 30µg of proteins were fractionated using 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The experimenters used GAPDH as a loading control. A gel documentation system was used for band identification, and computer software was used to quantify band intensities.
Strengths and Limitations
Vertical electrophoresis tanks are ideal for protein separation and identification. In vertical electrophoresis systems, the buffer can only migrate through the gel, allowing accurate voltage control during experiments. If the polyacrylamide gel with a small pore size is used, the vertical gel electrophoresis allows greater separation and resolution than horizontal systems. However, horizontal gel electrophoresis systems are sometimes preferred over vertical systems because of their ease of use.
- Always wear PPE like safety goggles while working with UV radiation.
- Do not leave the running vertical electrophoresis tank unattended.
- Do not touch any apparatus connected to the gel as the tubing can conduct current.
- If you use ethidium bromide as a stain, handle it with care. Work in a fume hood and wear proper PPE like gloves and lab coats. EtBr is a potential carcinogen, and exposure to it can be hazardous.
- Vertical electrophoresis tanks are primarily used for nucleic acid and protein sequencing via vertical gel electrophoresis.
- In a vertical electrophoresis system, the gel matrix is cast vertically in a discontinuous buffer system, and two chambers, i.e., a top chamber and a bottom chamber, are employed.
- As soon as the electric field is applied, a small amount of buffer migrates through the gel from the upper chamber towards the lower chamber.
- Vertical systems mostly use polyacrylamide gels with a smaller pore size (about 10-200nm).
- Vertical gel electrophoresis is ideal for the separation of proteins.
Iukalo, А. V. (2015). Identification of protein fractions of milk cows casein complex. The Ukrainian Biochemical Journal, (87,№ 4), 87-91.
Han, D., Wu, C., Xiong, Q., Zhou, L., & Tian, Y. (2015). Anti-inflammatory mechanism of bone marrow mesenchymal stem cell transplantation in rat model of spinal cord injury. Cell biochemistry and biophysics, 71(3), 1341-1347.
Model 1, Model 2