6% SDS-PAGE colored (green) gel ultra-fast preparation kit, 50 T
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SKU : G2060-50T
หมวดหมู่ : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
แบรนด์ : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
Fast-Cast Colorful (Green) Acrylamide Kit, 6% | G2060-50T | 50 T (1.0 mm Glass Plates) |
Product Description/Introduction
This kit provides a simple and rapid preparation of 6% PAGE coloured (green) gels, enabling the simultaneous preparation of both resolving and stacking gels, allowing the user to prepare protein gels quickly by simply preparing the gel preparation equipment. At the same time, the kit contains a green dye in the stacker B, which makes it possible to visualise the spiking process and greatly improves the spiking efficiency. The green dye is stable in the stacker and does not migrate to the resolver with electrophoresis, thus not affecting the electrophoresis and staining effect. It is also easy to identify and excise the stacker after electrophoresis is completed, without affecting subsequent experiments such as Western Blot. The gels prepared in this kit can also be used for non-denaturing PAGE gel electrophoresis.
This kit prepares 50 gels of regular size, the number of gels prepared depends on the thickness of the gel and the size of the gel.
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 | G2060-50T |
G2060-1 | Stacker A | 50 mL |
G2060-2 | Stacker B (Green) | 50 mL |
G2060-3 | 6% Resolver A | 125 mL |
G2060-4 | 6% Resolver B | 125 mL |
G5036-5ML | Modified coagulant | 1 mL*5 |
Manual | 1 pc | |
Assay Protocol/Procedures
1. Choose appropriate Concentration of PAGE resolver (separation gel) according to molecular weight of target protein, refer to the table below:
PAGE Separating Gel Concentration | Best separation range (kDa) (Tris-Glycine Buffer, G2018) | Best separation range (kDa) (SWE Fast High-Resolution Electrophoresis Buffer, G2081) |
6% | 50-250 kDa | 15-300 |
8% | 30-130 kDa | 10-250 |
10% | 20-100 kDa | 5-150 |
12% | 10-60 kDa | 3-100 |
15% | < 40 kDa | <60 |
2. According to the experimental requirements, mix solution A and B in equal proportion, add appropriate amount of modified coagulant, and prepare the resolving solution and stacking solution respectively. Different specifications and different thickness of the glass plate can be adjusted in equal proportions to the stacking solution and resolving solution preparation volume, to the commonly used specifications of 8.3 cm × 7.3 cm gel plate (single), for example, the recommended preparation system is as follows table:
Configuration Groups | Component | 0.75 mm Glass Plates | 1.0 mm Glass Plates | 1.5 mm Glass Plates |
Resolver | 6% Resolver A | 2 mL | 2.5 mL | 4 mL |
6% 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 (Green) | 1 mL | 1 mL | 1.5 mL | |
Modified coagulant | 12 μL | 12 μL | 18 μL |
3. 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, and then add the prepared stacker, insert the comb, and wait until it is solidified (about 10-15 min), and then can be used; or in the stacker can be added to join the prepared resolver immediately and slowly, uniformly added to the upper layer of the gel solution prepared, and insert the comb, and wait until it is solidified (about 10-15 min), and then can be used.
4. If the prepared gel cannot be used on the same day, it can be stored at 4°C for 1-2 days.
5. Recommended electrophoresis conditions:
a. Use SWE Fast High-Resolution Electrophoresis Buffer (G2081) for electrophoresis: 200-250 V, 25-35 min.
b. Use Tris-Glycine Electrophoresis Buffer (G2018) for electrophoresis: stacker 90 V, about 30 min (marker enters separation gel); resolver 150-180 V, about 60-90 min (can be adjusted according to actual situation).
Notes:
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!
