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G2064-50T 15% SDS-Page Colored Green Gel Ultra-Fast Preparation Kit

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G2064-50T 15% SDS-Page Colored Green Gel Ultra-Fast Preparation Kit

This kit provides a simple and fast 15% SDS-PAGE color (green) gel preparation method.The stacking gel reagent can prepare a colored (green) upper gel, so we can easily identify each well of the gel when loading the sample. After the electrophoresis, the dye does not move to the separation 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.

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Item code Description Volume
G2064-1 Stacker A 50mL
G2064-2 Stacker B (Green) 50mL
G2064-3 15% Resolver A 125mL
G2064-4 15% Resolver A 125mL
G2064-5 APS (ammonium persulfate) 180mg x 3 (Powder)

Technical Specification

Storage:

Store at 4ºC, the shelf life is 1 year.

Assay Protocol / Procedures:

1. Choose appropriate Concentration of acrylamide solution according to molecular weight of target protein;

Concentration of acrylamide

Separation range

6%

50-250 kDa

8%

30-130 kDa

10%

20-100 kDa

12%

10-60 kDa

15%

< 40 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 4ºC for 1-2 weeks, and APS powder can be stored at -20ºC.

3. Preparation of 15% resolving gel acrylamide solution:in a suitable container,add equal volumes of 15% Resolver A and 15% Resolver B;

15% 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)

15% Resolver A

2 mL x n

2.5 mL x n

4 mL x n

15% Resolver B

2 mL x n

2.5 mL x n

4 mL x n

10% APS

40 uL x n

50 uL x n

80 uL x n

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 add 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 x n

1 mL x n

1.5 mL x n

Stacker B

0.8 mL x n

1 mL x n

1.5 mL x n

10% APS

16 uL x n

20 uL x n

30 uL x n

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 4ºC for up to one week.

Note:

  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 colors (red, yellow, blue) of stacking solutions to distinguish different samples and different gel electrophoresis;
  5. Tris-Glycine buffer system is suitable (recommend our product: G2018-1 L).
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