Product Description

Polyacrylamide gel (PAGE gel) is often used for protein electrophoresis to realize protein separation. This kind of gel is generally composed of concentrated gel and separation gel. The former plays the role of concentrating protein samples, while the latter separates proteins of different sizes according to the concentration of acrylamide monomer and N,N-methylenebisacrylamide (methyleneacrylamide) crosslinking agent used in the gel.
This kit provides a variety of reagents required for the quick preparation of PAGE gel. Users only need to prepare their gel preparation equipment to prepare gel, which greatly simplifies the gel preparation process. The gel prepared by this kit can only be used for denaturing PAGE gel electrophoresis. This specification can prepare about 125 pieces of mini gel (calculated by 0.75 mm thick gel).

Product Features

1. Fast gel preparation speed - at least 2 min is required to prepare multiple gel, without calculating the required solution amount and dilution;
2. Color concentrated gel - the concentrated gel is colored, which provides the greatest convenience for sampling. The color formula contained does not affect subsequent applications such as electrophoresis, dyeing, and membrane transfer;
3. No odor - TEMED is not required to avoid odor;
4. High stability - the ammonium sulfate catalyst solution provided by the kit is an improved APS, and its stability and catalytic efficiency have been greatly improved.
5. New function - no need to close the separation gel, and directly add concentrated gel (read the gel preparation step).

Product Components

Component		20324ES62（125 gels）	Storage
20324-A	8 % - separation gel buffer	250 mL	4℃
20324-B	8 % - separation gel solution	250 mL	4℃
20324-C	8 % - color concentrated gel buffer	80 mL	4℃
20324-D	8% - concentrated gel solution	80 mL	4℃
20324-E	Prepare gel cup	3	4℃
20324-F	Improved ammonium persulfate solution	8 mL	-20℃

Shipping and Storage

The products are shipped with ice packs and can be stored at 4℃. Among them, 20324-F (Improved ammonium persulfate solution) can be stored at -20℃ for 12 months if it is not used for a long time.

Cautions

1. It is suggested that the voltage during electrophoresis should be between 100-120 V. If it is necessary to accelerate the electrophoresis speed, it can be increased to 150 V.  
2. Before gel filling, make sure to balance the gel solution to room temperature (for example, keep it for several minutes) to effectively avoid the formation of bubbles in the gel.
3. The amount of ammonium persulfate solution (improved type) is only for reference, and the actual amount can be increased or decreased according to personal experimental habits and experience. Adding more ammonium persulfate can accelerate the gel speed, and vice versa. The coagulation rate of PAGE gel is closely related to the temperature and the amount of ammonium persulfate.
4. An appropriate amount of TEMED substitutes have been added to this product. If it is necessary to further accelerate the gel speed, an appropriate amount of TEMED can be added as required before dispensing.
5. There is a significant positive correlation between gel solidification and temperature. Under the same conditions, the higher the temperature, the faster the solidification speed.
6. Color concentrated gel may produce a small amount of precipitation during storage, which is a normal phenomenon. Please feel free to use it.
7. The improved APS was stored at -20℃. For convenience, the APS that has been opened and is in use can be stored at 4℃.
8. For your safety and health, please wear lab coat and disposable gloves to operate.
9. For research use only.

Instructions for use

It is recommended not to configure too many pieces of gel at one time to avoid adding the upper solution too late (2-3 pieces are recommended at most)

1. Select the appropriate gel concentration according to the molecular weight of the target protein (refer to table 1)
2. Gel making process (take a piece of 0.75/1.0/1.5 mm Mini gel as example)
2.1 Take an equal volume of separation gel buffer and separation gel solution and mix them, i.e. take 2.0/2.7/ 4.0 mL of the two solutions respectively.
2.2 Add 35/45/70 μL improved APS to the mixed solution in step 1 (the amount of APS used can be reduced by half if the solidification is too fast) and fully mixed.
2.3 Inject the solution in step 2 into the gel making glass plate. Note that the concentrated gel shall be injected into the gel mold within 2 minutes after the separation gel is added, and the pouring of the concentrated gel shall be slow to prevent mixing of the concentrated gel and the separation gel. If the individual finds it difficult to configure, it can also be adapted to configure concentrated gel after sealing with ethanol.
2.4 Preparation of concentrated gel: take an equal volume of concentrated gel buffer and concentrated gel solution, i.e. take 0.5/0.75/ 1.0 mL of the two solutions respectively, and then add 10/13/18 μL improved APS and mixed well.
2.5 Inject it into the gel making glass plate and insert the comb teeth (do not use excessive force to insert the comb teeth gently).
2.6 After the concentrated gel is solidified for 15 min, the comb teeth can be removed and used for electrophoresis. Note: please try to use freshly prepared electrophoresis buffer.

Table 1  Reference separation range of SDS-PAGE gel with different concentration

SDS-PAGE gel concentration	Separation range (kDa)
6%	70-300
8%	30-200
10%	20-80
12.5%	15-60
15%	10-45