1.SDS-PAGE Gel Preparation Kit, (any concentration, indication dyes), 50T $150
Description
Product Information
Product Name | Cat.No. | Spec. |
SDS-PAGE Gel Preparation Kit | G2003-50T | 50 T |
Description
SDS-PAGE gel preparation kit contains all reagents required for gel preparation. Users only need to prepare a small amount of pure water and glue-making equipment (glue-making base, glass plate, comb, etc.), and the required gel can be prepared according to the instructions. This kit can prepare at least 50 gel pieces of conventional size.
Storage and Handling Conditions
Transport with wet ice; Store at 4℃ away from light, valid for 12 months.
Component
Component Number | Component | G2003-50T |
G2003-1 | 1.5 M Tris-HCl(pH 8.8) | 100 mL |
G2003-2 | 30% Acrylamide-Bisacrylamide (29:1) | 100 mL |
G2003-3 | 1.0 M Tris-HCl(pH 6.8) | 20 mL |
G2003-4 | 10% SDS | 5 mL |
G2003-5 | AP ( Ammonium Persulfate ) | 3×180 mg |
G2003-6 | TEMED | 500 μL |
Product Manual |
Assay Protocol
1. According to the molecular weight of the target protein and the selected electrophoresis buffer, select the appropriate concentration of SDS-PAGE separation glue, as shown in the table below:
SDS-PAGE Separation Gel Concentrations | Optimum separation range(kDa)
(Tris-Glycine electrophoresis buffer, G2018) |
Optimum separation range(kDa)
(SWE Fast High Resolution Electrophoresis Buffer, G2081) |
6% | 50-300 | 15-300 |
8% | 30-130 | 10-250 |
10% | 20-100 | 5-150 |
12% | 10-60 | 3-100 |
15% | <40 | <60 |
2. According to the molecular weight of the target protein, the concentration of the separation glue was selected. Taking the common specification of 8.3 cm×7.3 cm gel plate (single block) as an example, the glue preparation solution could be prepared according to the following table:
Separation Gel Concentrations (%) | 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 |
Total volume of glue required*(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) | 2.12 | 3.18 | 4.24 | 1.85 | 2.78 | 3.7 | 1.59 | 2.38 | 3.17 | 1.32 | 1.98 | 2.64 | 0.92 | 1.38 | 1.84 |
30% Acrylamide-Bisacrylamide (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 |
1.5 M Tris-HCl(pH 8.8)(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% SDS(μL) | 40 | 60 | 80 | 40 | 60 | 80 | 40 | 60 | 80 | 40 | 60 | 80 | 40 | 60 | 80 |
10% AP ( Ammonium Persulfate ) μL | 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 glue preparation solution does not include the volume of TEMED.
3. The preparation of concentrated glue can refer to the following table:
浓缩胶浓度(%) | 5% | |||
H2O(mL) | 1.93 | 2.95 | 3.86 | 5.79 |
30% Acrylamide-Bisacrylamide (29:1) (mL) | 0.5 | 0.75 | 1.0 | 1.5 |
1.0M Tris-HCl(pH 6.8)(mL) | 0.5 | 0.75 | 1.0 | 1.5 |
10% SDS(μL) | 40 | 60 | 80 | 120 |
10% AP ( Ammonium Persulfate )(μL) | 30 | 45 | 60 | 90 |
TEMED(μL) | 4 | 6 | 8 | 12 |
Total volume*(mL) | 3 | 4.5 | 6.0 | 9.0 |
* The total volume does not include the TEMED volume.
4. Recommended electrophoretic conditions:
a. Using SWE fast high-resolution electrophoresis buffer (G2081) for electrophoresis: 200-250 V constant pressure, 25-35 min to complete the electrophoresis;
b. Tris-Glycine electrophoresis buffer (G2018) was used for electrophoresis: the voltage of the upper glue was set at 90 V, and electrophoresis lasted for about 30 min (marker entered the separating glue); The adjustment voltage of the lower rubber is 150-180 V, about 60-90 min (can be adjusted according to the actual situation).
Note:
1. All reagents should be rewarmed to room temperature before use.
2. AP (ammonium persulfate) was prepared with pure water to 10% concentration, that is, 1800 μL of pure water was added to each 180 mg AP (ammonium persulfate) tube to dissolve. The AP solution prepared is recommended to be divided into small parts and stored at -20℃ for 3 months. If stored at 4℃, it will be valid for 1 week.
3. Tricine gel is recommended for protein separation smaller than 10 kDa; acrylamide gels may not be sufficient for separation.
4. 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.
5. The gel needs sufficient solidification time, and it is recommended that the gel be fully prepared and left to ensure that the gel is thorough.
6. When the temperature is low, 10%SDS solution has crystallization precipitation, and can be used after remelting at 37℃.
7. If you need a full set of reagents, you can buy other products of our company, such as SDS-PAGE gel preparation kit (G2003/G2037), super-fast color gel preparation kit series or super-fast color gel preparation kit series (G2041 / G2042 / G2043 G2044 / G2045 / G2060 / G2061 / G2062 / G2063 / G2064), And high resolution ultra-fast color gel preparation kit series (G2066/G2067/G2068, G2071/G2072/G2073), can meet different experimental needs.
8. For your safety and health, please wear a lab coat and disposable gloves when operating.
For Research Use Only!
Cat.No.
|
Product Name
|
Spec.
|
Operation
|
---|
G2002-100ML
|
RIPA Lysis Buffer (Strong)
|
100 mL
|
|
G2002-30ML
|
RIPA Lysis Buffer (Strong)
|
30 mL
|
|
G2003-50T
|
SDS-PAGE Gel Preparation Kit
|
50 T
|
|
G2004-100ML
|
30% Acrylamide-Bisacrylamide (29:1)
|
100 mL
|
|
G2006-250UL
|
50×Cocktail Protease Inhibitor
|
250 μL
|
|
G2007-1ML
|
Phosphoprotease Inhibitor
|
1 mL×2
|
|
G2008-1ML
|
PMSF (100mM)
|
1 mL
|
|
G2018-1L
|
Tris-Glycine SDS-PAGE Running Buffer (Powder)
|
1 L
|
|
G2033-100ML
|
RIPA Lysis Buffer (Weak)
|
100 mL
|
|
G2033-30ML
|
RIPA Lysis Buffer (Weak)
|
30 mL
|
|
G2053-100ML
|
1.5 M Tris-HCl (pH 8.8)
|
100 mL
|
|
G2054-100ML
|
1 M Tris-HCl (pH 6.8)
|
100 mL
|
|
G2055-5ML
|
10% SDS Solution
|
5 mL
|
|
G6019-9
|
Antibody Incubation Box (9 Grids)
|
9 grids
|
|
G6020-9
|
Antibody Incubation Box (9 Grids Light-Proof)
|
9 Grids Light-Proof
|
|
G6025-2
|
Antibody Incubation Box (2 Grids)
|
2 grids
|
|
G6026-4
|
Antibody Incubation Box (4 Grids)
|
4 grids
|