SDS-PAGE gel fast preparation kit, 50 T
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SKU : G2037-50T
Categories : Biochemicals , 1. Chemical and Reagents , Buffers , Servicebio ,
Brand : Servicebio
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Product Information
Product Name | Cat. No. | Spec. |
SDS-PAGE Fast Acrylamide Kit | G2037-50T | 50 T |
Product Description/Introduction
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 gel preparation equipment (gel base, glass plate, comb, etc.) and follow the instructions to produce the required gel in a short time. This kit can prepare at least 50 regular sized 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 | 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 | PAGE gel solidification catalyst | 1 mL |
G5036-5ML | Modified Coagulant | 1 mL×5 |
Manual | One copy | |
Assay Protocol/Procedures
1. Choose appropriate Concentration of acrylamide solution according to molecular weight of target protein;
SDS-PAGE separating gel concentration | The optimal Separation range (kDa) (Tris-Glycine running buffer, G2018) | The optimal Separation range (kDa) (SweRapid-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. The concentration of the separating gel was selected according to the molecular weight size of the target protein, take the gel plate (single piece) with common specifications of 8.3×7.3 cm as an example, the following table can be referred to for the preparation of the gel solution:
Concentration of separating gel(%) | 6% | 8% | 10% | 12% | 15% | ||||||||||
Glass plate thickness (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 | 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 |
Modified Coagulant (µL) | 40 | 60 | 80 | 40 | 60 | 80 | 40 | 60 | 80 | 40 | 60 | 80 | 40 | 60 | 80 |
PAGE gel solidification catalyst（µL） | 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 |
*The total volume of the gel solution does not include the volume of PAGE gel solidification catalyst.
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 |
Modified Coagulant/µL | 18 | 27 | 36 | 54 |
PAGE gel solidification catalyst/μL | 4 | 6 | 8 | 12 |
Total volume/mL | 3 | 4.5 | 6 | 9 |
*The total volume does not include the volume of PAGE gel solidification catalyst.
4. The recommended electrophoresis conditions:
a) Use SweRapid-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 is set at a voltage of 90 V, and electrophoresis is 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).
Note
1. Too large a temperature difference in the process of making glue heat production may produce bubbles, it is recommended that the glue is ready to return to room temperature, and then add the improved coagulant for gelation, which can effectively avoid the formation of small bubbles in the gelation process.
2. The modifiedhas better stability compared to ammonium persulfate (AP). Take out one for use, and store it at 4 after use for subsequent routine use, which can be stored for six months. If not used for a long time, please store it at -20 to avoid repeated freezing and thawing.
3. Tricine gel is recommended for protein separations smaller than 10 kDa; acrylamide gels may not be sufficient for separation.
4. Temperature has a significant impact on the gelation time of the adhesive. To ensure smooth experiment progress, generally, lower temperatures result in longer gelation times, and it may be necessary to increase the dosage of the modifiedcoagulant accordingly. On the other hand, higher temperatures lead to faster gelation, so the dosage of the modified coagulant can be reduced accordingly.
5. Adequate gelation time is needed, and it is recommended to allow the gel to sit undisturbed to ensure thorough gelation.
6. When the temperature is low, buffer solutions for separating gel and concentrated gel may crystallize due to the presence of SDS solution. They can be rewarmed at 37 and then used after melting.
7. If there is a need for super-fast and high-resolution gel preparation reagents, you can purchase our company's other products: Super Fast Colorful Gel Preparation Reagent Kit Series (G2041/G2042/G2043/G2044/G2045,G2060/G2061/G2062/G2063/G2064),and High-Resolution Super Fast Colorful Gel Preparation Reagent Kit Series (G2066/G2067/G2068, G2071/G2072/G2073), which can meet different experimental needs.
1. Please wear lab coat and disposable gloves during operation.
For Research Use Only!
