Gradient-like PAGE Gel Rapid Electrophoresis Kit

Product Introduction

This product is designed specifically for SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and includes a complete set of reagents for gel preparation as well as the necessary electrophoresis buffers. The resulting gels offer broad-spectrum separation capabilities (10–250 kDa) comparable to gradient gels, ensuring uniform distribution of low-, medium-, and high-molecular-weight proteins without the need for varying resolving gel concentrations; furthermore, electrophoresis can be completed in just 25 minutes at a constant voltage of 200 V.

The kit utilizes a pre-mixed formulation for both the stacking and resolving gels; polymerization is initiated simply by adding the modified coagulant. After pouring the resolving gel, the stacking gel can be poured immediately without waiting for the resolving gel to set, making the process quick and convenient. The prepared stacking gel is colored (red, blue, or green), ensuring that the sample wells are distinct and easy to identify for convenient sample loading.

This product comes with an improved polymerization accelerator that offers superior stability and catalytic efficiency; no additional TEMED is required during gel preparation. For convenience, the opened accelerator can be stored at 4°C for at least three months.

Item No.	Specification
FG-PG610	>125 gels (0.75 mm gel) or >90 gels (1.00 mm gel) or >60 gels (1.50 mm gel)

Component Name	Volume and quantity
Stacking gel solution (2×)	80 mL
Colored Stacking Gel Buffer (2×)	80 mL
Lower gel solution (2×)	250 mL
Lower gel buffer (2×)	250 mL
Improved coagulant	8 mL
FG-PG610 Specialized Electrophoresis Buffer Quick-Dissolving Granules	500 mL*40

Product Features

Wide separation range：The prepared gel exhibits broad-spectrum separation capabilities similar to the protein band patterns of gradient gels, eliminating the need to distinguish between lower-layer gels of different concentrations;

High electrophoresis speed：Electrophoresis can be completed in 25 minutes at a constant voltage of 200 V;

One-step potting：After casting the lower gel layer, the upper gel layer is injected directly without the need for a liquid overlay;

Simple and fast operation: No calculation of the required solution volume or dilution is necessary for gel preparation;

Colored Top Coat: Stacking gels in red, blue, and green can be prepared, facilitating sample loading and the differentiation of various gels;

Prevent unpleasant odors: Eliminates the need for TEMED, avoiding foul odors.

Adhesive Preparation Process

(Taking a 0.75/1.0/1.5 mm mini-gasket as an example)

1. Combine equal volumes of the resolving gel solution and resolving gel buffer (2.0, 2.7, or 4.0 mL of each) and mix well;

2. Mix equal volumes (0.5, 0.75, or 1.0 mL each) of the stacking gel solution and the colored stacking gel buffer.

Note: Due to the specific physicochemical properties of the dye, please shake well before use.

3. Add 40, 60, or 80 μL of the modified coagulant to the mixed solution from Step 1 and mix gently; then, pour the mixture into the glass gel-casting plates, ensuring the distance between the liquid surface and the top edge of the short glass plate exceeds the length of the comb teeth by 0.5 cm.

Note: ① This solution is in excess; do not inject the entire amount.

② After adding the modified coagulant, mix gently to prevent excessive oxygen from being incorporated into the gel solution, which would inhibit gel polymerization.

4. Add 10, 15, or 20 μL of the modified coagulant to the mixed solution from Step 2 and mix gently; without waiting for the lower gel layer to solidify, gently inject the mixed solution into the gel-casting glass plates and insert the comb.

Note: ① Pour the upper gel solution gently and slowly to avoid disrupting the lower gel layer;

② After adding the modified polymerization initiator, mix gently to prevent excessive oxygen incorporation, which could inhibit gel polymerization.

5. Once the gel has solidified (approximately 15 minutes), remove the comb, and it is ready for electrophoresis.

Note: ① Please use freshly prepared electrophoresis buffer whenever possible;

② After the gel solidifies, the interface between the upper and lower gel layers is slightly less uniform than that of gels prepared using traditional methods, but this does not affect subsequent electrophoresis.

Base coat formulation				Top-layer adhesive formulation
Gel thickness	Lower gel solution	Lower buffer	Improved coagulant	Gel thickness	Upper gel solution	Stacking gel buffer	Improved coagulant
0.75 mm	0.75 mm	0.75 mm	0.75 mm	0.75 mm	0.75 mm	0.75 mm	0.75 mm
1.00 mm	1.00 mm	1.00 mm	1.00 mm	1.00 mm	1.00 mm	1.00 mm	1.00 mm
2.0 mL	2.0 mL	2.0 mL	2.0 mL	2.0 mL	2.0 mL	2.0 mL	2.0 mL

Electrophoresis

1. Prepare the electrophoresis buffer.

a. Pour approximately 300 mL of distilled water into a beaker, slowly add one packet of PG610-specific electrophoresis buffer granules, and stir the solution until the granules are completely dissolved;

b. Add distilled water to the solution obtained in step (a) and adjust the volume to 500 mL to obtain 1× electrophoresis buffer.

2. Recommended electrophoresis conditions

Constant voltage of 150 V for approximately 35 min, or constant voltage of 200 V for approximately 25 min.

Precautions

1. The gel prepared with this product must be used with the dedicated PG610 electrophoresis buffer; do not substitute with other electrophoresis buffers, as doing so will prevent achieving optimal electrophoresis results.

2. When performing Western blot transfer, the transfer conditions for standard 10% PAGE gels may be used for gels prepared with this product.

3. If better separation of low-molecular-weight protein bands is required, simply adjust the ratio of the resolving gel solution to the resolving gel buffer to 1.1:1 when preparing the gel; conversely, for better separation of high-molecular-weight protein bands, adjust the ratio to 0.9:1.

4. The usage amount of the modified coagulant serves only as a reference; the actual quantity may be adjusted based on individual experimental habits and experience. The gelation rate is significantly positively correlated with temperature. Under identical conditions, a higher temperature results in a faster gelation rate; therefore, it is advisable to reduce the amount of modified coagulant if the room temperature is excessively high. Conversely, if the room temperature is low, the gelation time may be appropriately extended.

5. Equilibrating the gel solution and buffer to room temperature (e.g., by letting them sit for a few minutes) before preparing the gel can effectively prevent the formation of air bubbles within the gel.

6. For your safety and health, please wear a lab coat and disposable gloves while performing the procedure.

7. This product is for research use only.