SDS-PAGE Fast Acrylamide Kit
Cat. No.: SB-G2037-50T
Size: 50T
Description
This kit provides a simple and fast SDS-PAGE gel preparation solutions. It contains all the reagents needed for gel preparation, users only need to prepare a small amount of pure water and glue making equipment (glue base, glass plate, comb, etc. ), according to the instructions to produce the required gel in a short time. This kit can prepare at least 50 pieces of conventionally sized gel.
Storage and transportation
Shipment by ice bag, Store at 4 ℃ , the shelf life is 1 year .
Product Contents
| Component Number |
Component |
G2037-50T |
| G2037-1 |
30% acrylamide (29:1) |
100 mL |
| G2037-2 |
4 × Tris-SDS Lower Separating Gel Solution |
100 mL |
| G2037-3 |
4 × Tris-SDS Upper Stacking Gel Solution |
30 mL |
| G2037-4 |
AP (ammonium persulfate) |
3 × 180 mg (Powder) |
| G2037-5 |
TEMED |
500 μL |
| Product Manual |
1 copy |
Assay Protocol / Procedures
1. Choose appropriate Concentration of acrylamide solution according to molecular weight of target protein ;
| Concentration of Lower Acrylamide |
The optimal separation range
(Tris-Glycine running buffer, G2018) |
The optimal separation range
(Swe Rapid-High Resolution running buffer, G2018) |
| 6% |
50-300 kDa |
15-300 kDa |
| 8% |
30-130 kDa |
10-250 kDa |
| 10% |
20-100 kDa |
5-150 kDa |
| 12% |
10-60 kDa |
3-100 kDa |
| 15% |
< 40 kDa |
< 60 kDa |
2. Take the gel plate (single piece) with common specifications of 8.3 × 7.3 cm as an example, the Lower acrylamide solution can be formulated by referring to the following table:
| Concentration of lower acrylamide(%) |
6% |
8% |
10% |
12% |
15% |
| Glass plate thickness |
0.75 mm |
1.0 mm |
1.5 mm |
0.75 mm |
1.0 mm |
1.5 mm |
0.75 mm |
1.0 mm |
1.5 mm |
0.75 mm |
1.0 mm |
1.5 mm |
0.75 mm |
1.0 mm |
1.5 mm |
| The total volume of gel solution* (mL) |
4.0 |
6.0 |
8.0 |
4.0 |
6.0 |
8.0 |
4.0 |
6.0 |
8.0 |
4.0 |
6.0 |
8.0 |
4.0 |
6.0 |
8.0 |
| H2O（mL) |
2.16 |
3.24 |
4.32 |
1.89 |
2.84 |
3.78 |
1.63 |
2.44 |
3.25 |
1.36 |
2.04 |
2.72 |
0.96 |
1.44 |
1.92 |
| 30% acrylamide(29:1) (mL) |
0.8 |
1.2 |
1.6 |
1.07 |
1.6 |
2.14 |
1.33 |
2.0 |
2.67 |
1.6 |
2.4 |
3.2 |
2.0 |
3.0 |
4.0 |
| 4 x Tris-SDS Lower Separating Gel Solution (mL) |
1.0 |
1.5 |
2.0 |
1.0 |
1.5 |
2.0 |
1.0 |
1.5 |
2.0 |
1.0 |
1.5 |
2.0 |
1.0 |
1.5 |
2.0 |
| 10% AP (ammonium persulfate) (uL) |
40 |
60 |
80 |
40 |
60 |
80 |
40 |
60 |
80 |
40 |
60 |
80 |
40 |
60 |
80 |
| TEMED（μL） |
2.0 |
3.0 |
4.0 |
2.0 |
3.0 |
4.0 |
2.0 |
3.0 |
4.0 |
2.0 |
3.0 |
4.0 |
2.0 |
3.0 |
4.0 |
*The total volume of the gel solution does not include the volume of TEMED.
3. Prepare the Upper Stacking Gel by referring to the following table:
| Concentration of Upper Stacking Gel |
5% |
| H2O/mL |
1.75 |
2.63 |
3.5 |
5.25 |
| 30% acrylamide (29:1)/mL |
0.5 |
0.75 |
1 |
1.5 |
| 4 x Tris-SDS Lower Separating Gel Solution/mL |
0.75 |
1.13 |
1.5 |
2.25 |
| 10% AP (ammonium persulfate)/uL |
30 |
45 |
60 |
90 |
| TEMED/μL |
4 |
6 |
8 |
12 |
| Total volume/mL |
3 |
4.5 |
6 |
9 |
*The total volume does not include the volume of TEMED.
4. The recommended electrophoresis conditions:
A. Use Swe Rapid-High Resolution running buffer ( G2018 ) at constant voltage of 200-250 V for 25-35 min .
B. Or use Tris-Glycine running buffer (G2018). The upper gel was set at a voltage of 90 V, and electrophoresis was performed for about 30 min (marker entered the separation gel). The voltage of the lower gel is adjusted to 150-180 V, about 60-90 min (Adjust according to the actual situation).
1. All reagents should be rewarmed to room temperature before use .
2. 10% AP (ammonium persulfate) is prepared by adding 1800 μ L of pure water to each 180 mg AP (ammonium persulfate) tube in a 10% concentration. It is recommended that the AP prepared in solution be divided into small parts and stored at -20 ℃ for 3 months. If stored at 4 ℃ , valid for 1 week.
3. Temperature will have an effect on the solidification time of glue, generally the lower the temperature, the longer the solidification time; the higher the temperature, the faster the gel .
4. PAGE Gels need sufficient solidification time, it is recommended to stand fully after gelation to ensure that the gel is thorough
5. When the temperature is low, the separated glue buffer and the concentrated glue buffer will crystallize out due to the SDS solution, which can be used after rewarming at 37 ℃ .
6. Please wear lab coat and disposable gloves during operation . .
