SKU : G2159-50T
หมวดหมู่ : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
แบรนด์ : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
SDS-PAGE Colour (Red) Gel Cast Kit (Low Molecular Protein Separation, Tricine System) | G2159-50T | 50 T |
Product Description/Introduction
This kit provides a simple and rapid preparation reagent for SDS-PAGE colored (red) gel (low molecular weight protein separation, Tricine system), capable of simultaneously preparing stacking, spacer and resolving gels. Users only need to prepare the gel casting apparatus to rapidly prepare protein gels. Additionally, the solution B in the stacking 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 stacking gel without migrating to the resolving gel during electrophoresis, thus not affecting electrophoresis and staining results. It is also easy to identify and excise the stacking gel after electrophoresis completion, without impacting subsequent experiments such as Western Blot. Gels prepared with this kit are specifically designed for the separation of low molecular weight proteins (20 kDa). After formulation, the concentration of the stacking gel is 4%, the concentration of the spacer/resolving gel, spacer and stacking gel is 10%, and the concentration of the resolving gel is 16%.
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 |
G2159-1 | Stacker A | 50 mL |
G2159-2 | Stacker B (Red) | 50 mL |
G2159-3 | Spacer A | 45 mL |
G2159-4 | Resolver A | 90 mL |
G2159-5 | Spacer/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 stacker and resolver 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 | 1.2 mL | 1.5 mL | 2.4 mL |
Spacer/Resolver B | 1.2 mL | 1.5 mL | 2.4mL | |
Modified Coagulant | 14 μL | 18 μL | 28 μL | |
Spacer | Spacer A | 0.6 mL | 0.75 mL | 1.2 mL |
Spacer/Resolver B | 0.6 mL | 0.75 mL | 1.2mL | |
Modified Coagulant | 7 μL | 9 μL | 14 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 spacer, then seal the surface of the spacer gel with distilled water or ethanol, wait for the middle gel to solidify sufficiently (about 10-15 minutes), discard the water or ethanol, and use filter paper to suck up the residual water or ethanol.
3. 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 spacer, slowly and evenly add the prepared stacker, insert the comb, and allow it to solidify (about 10-15 minutes), and then can be used.
4. Recommended electrophoresis conditions:
Perform ice bath electrophoresis using Tricine SDS-PAGE Running Buffer (Powder) (G2142): 100 V, 180-240 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 recommended formulation system table is only applicable to the electrophoresis apparatus produced by our company. The volumes of spacer and resolving gels required for different manufacturers' electrophoresis apparatus may vary slightly. Please make appropriate adjustments. Generally, the volume of spacer gel : resolving gel is 1:2, and ensure that the comb does not touch the spacer gel after insertion. The optimal distance from the bottom of the comb teeth to the spacer gel is about 2 cm.
4. The prepared gel should be used on the same day and should not be stored at 4°C for later use.
5. 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.
6. There is monomer acrylamide in the premixed solution, which is harmful to the human body. Please pay attention to protective measures during operation.
7. 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.
8. After adding the modified coagulant, cast the gel as soon as possible and do not leave it for a long time.
9. 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.
10. 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.
11. We also provide other colored (green, yellow, blue) stacker to distinguish different samples for gel electrophoresis.
12. For your safety and health, please wear laboratory clothes and disposable gloves during operation.
For Research Use Only!
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