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This kit provides a simple and fast 15% SDS-PAGE color (green) gel preparation reagent, which can prepare the lower gel (ie separation gel) and the upper gel (also called concentrated gel, accumulated gel) at the same time. The user only need to prepare gel-making equipment to quickly prepare protein gels. At the same time, the upper gel solution B in the kit contains a green staining solution, which makes the protein spotting hole clear, realizes the visualization of the sample addition process, and greatly improves the sample addition efficiency. The green staining solution can stably exist in the upper gel, and will not migrate to the lower gel with electrophoresis, and will not affect the electrophoresis and staining effect. After the electrophoresis is completed, the upper gel can be easily identified and removed without affecting subsequent Western Blot and other experiments.
This kit can prepare 50 gels of conventional size, the specific number of gel blocks is related to the thickness of the gel and the size of the gel.
Storage and transportation
Wet ice Transportation ;
Stored at 4°C and away from light, valid for one year;
|G2064-1||Upper gel solution A||50mL|
|G2064-2||Upper gel solution B (green)||50mL|
|G2064-3||15% lower gel solution A||125mL|
|G2064-4||15% lower gel solution B||125mL|
|G2064-5||AP (Ammonium Persulfate)||1g(Powder)|
|SDS-PAGE separation gel concentration||Best separation range (kDa)|
2.Prepare 10% AP solution: Get 0.1 g of AP (ammonium persulfate) powder and dissolve it completely with 1 mL of water. The solution is valid for 1-2 weeks when stored at 4°C, and valid for 3 months when stored at -20°C.
3.According to the experimental requirements, mix solution A and B in equal proportions, add an appropriate amount of 10% AP solution, and prepare the lower gel solution and the upper layer gel solution respectively. For glass plates of different specifications and different thicknesses, the preparation volume of the upper gel and the lower gel solution can be adjusted in equal proportions. Taking the commonly used 8.3 cm×7.3 cm gel plate as an example, the recommended preparation system is as follows:
|Group||Component||Single piece of gel (1.0 mm thickness)||Two pieces of gel (1.0 mm thickness)|
Lower gel solution
|15% lower gel solution A||2.5mL||5mL|
|15% lower gel solution B||2.5mL||5mL|
|10% AP solution||50μL||100μL|
Upper gel solution
|Upper gel solution A||1mL||2mL|
|Upper gel solution B (green)||1mL||2mL|
|10% AP solution||20μL||40μL|
4. Prepare the gel tool and pour the prepared lower layer gel solution, and immediately add the prepared upper layer gel solution slowly and evenly (distilled water can also be used to seal the lower layer gel liquid surface, 15-30 minutes later until the lower layer gel solidifies, add the prepared upper layer gel solution after it is full), insert the comb, and wait for it to solidify (about 15-30 min) before usage.
5. If the prepared gel cannot be used on the same day, it can be stored at 4°C for 1-2 days before usage.
1. There is monomer acrylmide in the premix, which is harmful to the human body. Please take protective measures during operation.
2. In order to ensure that the sample enters the separation gel at the same time, when directly pouring the top gel, the prepared top gel solution should be added slowly and evenly.
3. Pour the gel as soon as possible after adding AP, do not leave it for a long time.
4. The solidification of the gel has a significant positive correlation with the temperature. Under the same conditions, the higher the temperature, the faster the solidification rate.
5.This product has been added with an appropriate amount of TEMED substitutes. If you need to further accelerate the gel speed, you can add an appropriate amount of TEMED as needed before the gel is prepared, and at the same time increase the AP usage by 0.5 times.
6. The company also provides other color (red, yellow, blue) top gel solutions to distinguish different samples and different gel electrophoresis.
7. The running buffer is suitable for the Tris-Glycine buffer system (recommended our product: G2018 running buffer).