How to use the principle of Hengaode instrument analytical electrophoresis instrument
How to use the principle of Hengaode instrument analytical electrophoresis instrument
The electrophoresis instrument was developed in 1937, and commonly used supporting objects include filter paper and acetate film. Electrophoresis is an indispensable and important analytical method for molecular biology research. Electrophoresis is generally divided into two categories: free interface electrophoresis and zone electrophoresis. Free interface electrophoresis does not require supports, such as isoelectric focusing electrophoresis, constant velocity electrophoresis, density gradient electrophoresis, and microelectrophoresis. .
Instrument principle
Zone electrophoresis requires various types of substances as supports. Commonly used supports include filter paper, cellulose acetate film, non-gel support, gel support and silica gel-G thin layer, etc., in the field of molecular biology The most commonly used is agarose gel electrophoresis. The so-called electrophoresis refers to the movement of charged particles in the electric field. Different substances have different speeds in the electric field due to the different charges and molecular weights. According to this feature, the electrophoresis method can be used to qualitatively or quantitatively analyze different substances. , Or a certain mixture for component analysis or single component extraction preparation, which has extremely important significance in clinical testing or experimental research. The electrophoresis instrument is designed and manufactured based on the above principles.
Instructions
1. First connect the two electrodes of the electrophoresis tank to the DC output terminal of the electrophoresis instrument with wires, pay attention not to reverse the polarity.
2. The power switch of the electrophoresis instrument is turned to the off position, the voltage knob is turned to the minimum, and the voltage stabilizing current mode and the voltage and current range are selected according to the work needs.
3. Turn on the power, slowly rotate the voltage adjustment knob until the desired voltage is reached, and set the electrophoresis termination time, at which time electrophoresis begins.
4. After the work is completed, the knobs and switches should be turned to the zero position or turned off, and the electrophoresis plug should be pulled out.
Precautions
1. After the electrophoresis instrument is energized and enters the working state, it is forbidden for the human body to contact the electrodes, electrophoresis objects and other parts that may be charged, nor to take things into the electrophoresis tank. If necessary, power off first to avoid electric shock. At the same time, the instrument must have a good ground terminal to prevent leakage.
2. After the instrument is powered on, do not temporarily add or remove the output lead plug to prevent short circuit phenomenon. Although a fuse is attached inside the instrument, the short circuit phenomenon may still cause damage to the instrument.
3. Because the resistance value of different media supports is different, the amount of current passed during electrophoresis is also different, the swimming speed and the time required for swimming to the end are also different, so electrophoresis of different media supports should not be on the same electrophoresis instrument at the same time. get on.
4. When the total current does not exceed the rated current of the instrument (maximum current range), it can be used in multiple slots, but be careful not to overload, otherwise it will easily affect the life of the instrument.
5. In some special cases, when the electrophoresis input condition of the instrument needs to be checked, it is allowed to start with no load under steady state, but it must be connected to the load before starting under steady state, otherwise the pointer of the voltmeter will jump greatly, which is easy to cause Unnecessary man-made machine damage.
6. If abnormal phenomena such as loud noise, discharge or unusual smell are found during use, the power supply shall be cut off immediately and repaired to avoid accidents.
R & D background
In 1937, the Swedish biochemist Tiselius gathered the predecessors for more than one hundred years to explore the phenomenon of electrophoresis, invented the Tiselius electrophoresis instrument, and established a free interface electrophoresis method for protein research on this basis. This method was used to prove for the first time that human serum is white. The protein (A), α, β, γ globulin, and therefore won the Ago Award in 1948. Subsequent developments in electrophoresis technology progressed by leaps and bounds. In 1949, RicketlsMarrack et al. Proved that human serum proteins can be divided into albumin, α1, α2, β, and γ globulin by electrophoretic separation. In 1957, Reiner treated five groups of human serum Protein fractionation was quantitatively analyzed.
However, free interface electrophoresis does not have a fixed support medium, and diffusion and convection have a strong effect, which affects the separation effect. Therefore, solid-phase support medium electrophoresis appeared one after another in the 1950s. The initial supporting media were filter paper and cellulose acetate membrane, and these media are currently used less in the laboratory. For a long period of time, small molecular substances such as amino acids, peptides, sugars, etc. are usually separated and analyzed by filter paper, cellulose or silica gel thin-layer plates, but currently more sensitive techniques such as high performance liquid chromatography are generally used Method (HPLC), etc. to analyze. For complex biological macromolecules, using filter paper, silica gel or cellulose acetate membrane as the supporting medium for electrophoresis, the separation effect is not ideal. So in 1959, Raymond and Weintraub, Davis and Ornstein successively used synthetic gel as a supporting medium to establish polyacrylamide gel electrophoresis, which greatly improved the resolution and separation effect of electrophoresis, and enhanced the development, penetration and The ability to combine with other technologies. As a result, all kinds of electrophoresis technologies and electrophoresis materials have sprung up and competed for prosperity, which has become a large technology with a wide variety, wide application, basic and cutting-edge technology in contemporary experimental science and technology.
According to whether the support medium is used in electrophoresis, it is divided into free electrophoresis and zone electrophoresis.
Free electrophoresis does not use supporting media, electrophoresis is performed in solution. This type of electrophoresis is divided into non-free interface electrophoresis and free interface electrophoresis. Non-free interface electrophoresis refers to that all charged particles (such as various cells) suspended in a solution move completely after being energized without an interface, such as microelectrophoresis. In free interface electrophoresis, the separated substances are concentrated in a certain layer, forming their own interfaces for qualitative or quantitative analysis. Free interface electrophoresis requires expensive and precise current instruments, and is only used in a few special electrophoreses such as isoelectric focusing electrophoresis and isokinetic electrophoresis.
Supporting media are used for zone electrophoresis, which can be divided into filter paper electrophoresis, acetate membrane electrophoresis, thin layer electrophoresis and gel electrophoresis according to different support media. In addition, it can be divided into horizontal plate electrophoresis, vertical plate electrophoresis, vertical disc electrophoresis, capillary electrophoresis, bridge electrophoresis, continuous flow electrophoresis, etc. according to the different types of devices supporting the medium.
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