BTT Fast-Cast Colorful (Green) Acrylamide Kit, 8%
Cat. No.: SB-G2071-50T
Size: 50T
Description
This kit provides a simple and fast 8% SDS-PAGE color ( green) gel preparation solutions . It can cast resolving and stacking gels at the same time and start running your gels in as short as 30 min .Also, it works for MOPS running buffer system, which can conduct higher resolution and sharper band than Laemmli SDS-PAGE system.
The stacking gel reagent is dyed green, so we can easily identify each well of the gel when loading the sample. After the electrophoresis, the dye does not move to the resolving gel, so it will not affect subsequent experiments.
In addition, due to the neurotoxicity of TEMED , safer alternatives have been used to provide a safer experience for operators.
Storage and transportation
25 X SDS Running buffer can be stored at room temperature;
other reagents can be store at 4℃;
The shelf life is 1 year.
Product Contents
| Component Number |
Component |
G2071 |
| G2071-1 |
BTT Stacker A |
50 mL |
| G2071-2 |
BTT Stacker B（Green） |
50 mL |
| G2071-3 |
8% BTT Resolver A |
125 mL |
| G2071-4 |
8% BTT Resolver B |
125 mL |
| G2071-5 |
APS (Ammonium Persulfate) |
180 mg x 3 ( Powder ) |
| G2071-6 |
25 X SDS Running buffer |
500 mL |
Assay Protocol / Procedures
1. Choose appropriate Concentration of acrylamide solution according to molecular weight of target protein ;
| Concentration of Acrylamide and Separation Range |
| Concentration of Acrylamide |
Separation range |
| 8% |
≥35 kDa |
| 10% |
10-250 kDa |
| 12% |
10-100 kDa |
2. Prepare 10% APS solution :100 mg of APS powder is dissolved in 1.0 mL of ultrapure water to obtain a 10% APS solution;
Note: 10% APS solution can be stored at 4 ° C for 1-2 weeks, and APS powder can be stored at -20 ° C .
3. Preparation of 8% resolving gel acrylamide solution : in a suitable container, add equal volumes of 8% Resolver A and 8% Resolver B
| 8% Resolving Gel Preparation Volume |
| |
0.75 mm Glass Plates (n=gels) |
1.0 mm Glass Plates (n=gels) |
1.5 mm Glass Plates (n=gels) |
| 8% Resolver A |
2 mL xn |
2.5 mL xn |
4 mL xn |
| 8% Resolver B |
2 mL xn |
2.5 mL xn |
4 mL xn |
| 10% APS |
40 uL xn |
50 uL xn |
80 uL xn |
4. Add the required volume of freshly made 10% APS to the combined resolver solution and mix well. Use an appropriate pipet to steadily dispense the solution into the cassette. Do not let bubbles form or solution mix with air. Fill the cassette to 0.5 -1 cm below the bottom of the teeth on the comb. Immediately and slowly add pure water or ethanol to the cassette. Allow the resolver to polymerize for 15-30 min, then pour water or ethanol out ;
Alternative : pour the stacking solution instead of water or alcohol as directed in the next steps.
5. Prepare stacking gel acrylamide solution by combining equal volumes of stacker A and B solution , then a dd required volume of freshly made 10% APS to the combined stacker solution and mix well .Pipet solution down the middle of the cassette, filling to the top of the short plates. Apply slowly and steadily to prevent mixing with the resolving solution. Align and insert the comb in the cassette carefully to prevent air from being trapped under the comb teeth;
6. After adding APS to the stacking solution mixture, casting should be started immediately and combs inserted within 8 min;
| Stacker Gel Preparation Volume |
| |
0.75 mm Glass Plates (n=gels) |
1.0 mm Glass Plates (n=gels) |
1.5 mm Glass Plates (n=gels) |
| Stacker A |
0.8 mL xn |
1 mL xn |
1.5 mL xn |
| Stacker B |
0.8 mL xn |
1 mL xn |
1.5 mL xn |
| 10% AP |
16 uL xn |
20 uL xn |
30 uL xn |
7. Allow the gel to polymerize for 15-30 min before electrophoresis;
8. The cast gels can be covered with a paper towel wetted with deionized water and stored in a zipper sealed plastic bag at 4 ℃ for up to one week .
9. Dilute 25 x SDS running buffer to 1 x SDS running buffer before electrophoresis.
1. There is monomer acrylamide in the premix, which is harmful to human body. Please pay attention to protective measures during operation
2. Pour the gel as soon as possible after adding APS , do not leave it for a long time ;
3. There is a significant positive correlation between the solidification and the temperature of the gel . Under the same conditions, the higher the temperature, the faster the solidification rate.
4. We also provide other color s (green, yellow, blue) of stacking solutions to distinguish different samples and different gel electrophoresis ; 5. It is recommended to use the 25 x running buffer provided in the kit instead of Tris-Glycine running buffer.
