SKU : G2155-50T
Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
PAGE Colour (Red) Gel Cast Kit (High Molecular Protein Separation) | G2155-50T | 50 T |
Product Description/Introduction
This kit provides a simple and rapid preparation reagent for PAGE colored (red) gel Cast Kit capable of simultaneously preparing stacking, resolving, and upper gels. Users only need to prepare the gel casting apparatus to rapidly prepare protein gels. Additionally, the solution B in the stacker gel of the kit contains a red dye, allowing for clear visualization of protein sample wells, greatly enhancing the efficiency of sample loading. This red dye can stably exist in the stacker gel without migrating to the resolver gel during electrophoresis, thus not affecting electrophoresis and staining results. It is also easy to identify and excise the stacker gel after electrophoresis completion, without impacting subsequent experiments such as Western Blot. The gel prepared in this kit does not contain SDS and can be used for denaturing or non-denaturing PAGE gel electrophoresis, which is specially used for the separation of large molecular proteins (300 kDa); the concentration of the stacker gel is 3%, and the concentration of the resolver gel is 7% after the formulation.
Storage and Shipping Conditions
Ship with wet ice; Modified coagulant stored at -20, others stored at 2-8 away from light, valid for 12 months.
Product Components
Component number | Component | G2159-50T |
G2155-1 | Stacker A | 50 mL |
G2155-2 | Stacker B (Red) | 50 mL |
G2155-3 | Resolver A | 125 mL |
G2155-4 | Resolver B | 125 mL |
G5036-5ML | Modified Coagulant | 1 mL×5 |
User Manual | 1 pc | |
Assay Protocol/Procedures
1. According to the experimental requirements, mix solutions A and B in proportion, add an appropriate amount of modified coagulant (see table below), and prepare the resovler and stacker soultions separately; different specifications and thicknesses of glass plates can adjust the preparation volume of the stacker and resolver solutions in proportion. Taking the commonly used 8.3 cm × 7.3 cm gel plate (single piece) as an example, the recommended preparation system is as follows in the table:
Configuration Groups | Components | 0.75 mm Glass Plates | 1.0 mm Glass Plates | 1.5 mm Glass Plates |
Resolver | Resolver A | 2 mL | 2.5 mL | 4 mL |
Resolver B | 2 mL | 2.5 mL | 4 mL | |
Modified Coagulant | 24 μL | 30 μL | 48 μL | |
Stacker | Stacker A | 1 mL | 1 mL | 1.5 mL |
Stacker B (Red) | 1 mL | 1 mL | 1.5 mL | |
Modified Coagulant | 12 μL | 12 μL | 18 μL |
2. After assembling the gel tool, first add the prepared resolver, then use distilled water or ethanol and other solutions to seal the surface of the resolver, wait until the resolver to solidify sufficiently (about 10-15 min), discard the water or ethanol, and use the filter paper to suck up the residual water or ethanol.Add the prepared stacker, insert the comb, and allow it to solidify (about 10-15 minutes) ,and then can be used. Alternatively, after adding the prepared resolver, slowly and evenly add the prepared stacker, insert the comb, and allow it to solidify (about 10-15 minutes) ,and then can be used.
3. If the prepared gel cannot be used on the same day, it can be stored at 4°C for 1-2 days.
4. Recommended electrophoresis conditions:
Perform electrophoresis using Tris-Acetate SDS-PAGE Running Buffer（Powder） (G2141): 100 V, 60-90 min.
Note
1. Too large a temperature difference in the process of making gel heat production may produce bubbles, it is recommended that A, B premixed solution is ready to return to room temperature, and then add modified coagulant for gelation, which can effectively avoid the formation of small bubbles in the gelation process.
2. The modified coagulant has better stability than ammonium persulfate (AP). After use, take out one and store it at 4°C for subsequent regular use. It can be stored for six months. If not used for a long time, please store it at -20°C to avoid repeated freezing and thawing.
3. The stacker B may produce flocculent precipitation after being left still for a long time. Please gently shake it before use to mix it evenly.
4. There is monomer acrylamide in the premixed solution, which is harmful to the human body. Please pay attention to protective measures during operation.
5. To ensure that the sample enters the separating gel at the same time, when directly casting the stacker, slowly and evenly add the prepared stacker.
6. After adding the modified coagulant, cast the gel as soon as possible and do not leave it for a long time.
7. Temperature has a significant effect on the solidification time of the gel. To ensure the smooth progress of the experiment, the lower the temperature, the longer the solidification time, and the amount of improved coagulant should be appropriately increased. The higher the temperature, the faster the gel solidifies, and the amount of improved coagulant can be appropriately reduced.
8. If you need to further accelerate the gel speed, add an appropriate amount of TEMED before casting the gel and increase the amount of modified coagulant by 0.5 times as needed.
9. We also provide other colored (green, yellow, blue) stacker to distinguish different samples for gel electrophoresis.
10. For your safety and health, please wear laboratory clothes and disposable gloves during operation.
For Research Use Only!
