Description
Gradient SDS-PAGE gel preparation kit
GenieBlot One-Step PAGE Gradient Gel Fast Preparation Kit is a rapid casting system for hand-pouring gradient SDS-PAGE gels in a single step. It gives reproducible separation across a wide molecular-weight range without dedicated gradient-mixing equipment, making it ideal for labs that prefer freshly cast gels with consistent resolution.
The premixed formulation lets you pour the stacking gel directly onto the resolving gel, with no sealing layer. Just mix the two solutions, add the polymerisation initiator (APS), and the gel sets within 10 min at room temperature. The brightly coloured stacking gel makes sample loading easy. The 4%–20% gradient separates proteins from 10 to 250 kDa with improved band resolution in standard Tris-Glycine running buffer, and electrophoresis is complete in 35–50 min at 150–200 V. Gels can be used for denaturing or non-denaturing PAGE and Western blotting.
Each kit makes 125 gels of 0.75 mm thickness. For research use only.
Key Features
| Wide separation range | Gradient separation of proteins from 10–250 kDa with uniform, clear bands. | |
| No special buffer | Compatible with standard Tris-Glycine electrophoresis buffer. | |
| Fast electrophoresis | Complete separation in 35–50 min at a constant 150–200 V. | |
| Rapid gel preparation | Mix two solutions, add initiator and the gel forms within 10 min. | |
| Easy loading | A coloured stacking gel makes wells easy to see. |
Specifications
| Application | Casting polyacrylamide gels for denaturing (SDS-PAGE) or non-denaturing PAGE |
| Gradient | 4%–20% |
| Separation range | 10–250 kDa |
| Gelation time | Within 10 min at room temperature (25°C) |
| Run conditions | 150–200 V for 35–50 min |
| Running buffer | Standard Tris-Glycine |
| Shelf life | 2 years |
Components
| Components | 125 gels/0.75 mm |
| Stacker A | 80 ml |
| Stacker B | 80 ml |
| Resolver A | 250 ml |
| Resolver B | 250 ml |
| APS | 12 ml |
Storage
Store at 2 ~ 8°C.
Volumes per mini gel by gel thickness. The steps below use a 1.0 mm mini gel as an example.
| Gel thickness | Resolver A | Resolver B | APS (resolving) | Stacker A | Stacker B | APS (stacking) |
| 0.75 mm | 2.0 ml | 2.0 ml | 40 μl | 0.5 ml | 0.5 ml | 10 μl |
| 1.0 mm | 2.7 ml | 2.7 ml | 60 μl | 0.75 ml | 0.75 ml | 15 μl |
| 1.5 mm | 4.0 ml | 4.0 ml | 80 μl | 1.0 ml | 1.0 ml | 20 μl |
- Mix the reagents
- Mix each component by inverting 6–8 times before use.
- Prepare the resolving (lower) gel
- Mix equal volumes of Resolver A and Resolver B (2.7 ml each).
- Prepare the stacking (upper) gel
- Mix equal volumes of Stacker A and Stacker B (0.75 ml each).
- Add APS
- Add 60 μl of APS to the resolving gel mix and 15 μl of APS to the stacking gel mix; mix each immediately by inversion.
- Mix without vortexing and avoid introducing bubbles.
- Pour the gel
- Quickly pour the resolving gel between the glass plates until the surface is about 1.0 cm below the top of the short plate.
- Without waiting for it to set, gently pour the stacking gel on top (within 2 min, so the lower gel surface stays even). Run it down the long glass plate to avoid disturbing the lower gel.
- Gently insert the comb. At room temperature (25°C) the gel sets within 10 min.
- Run the gel
- Remove the comb after gelation; the gel is ready to use immediately.
- Run at 150–200 V for 35–50 min, stopping when the bromophenol blue front reaches the bottom edge.
Q1: Do I need a special running buffer?
- No. The gels are compatible with standard Tris-Glycine electrophoresis buffer, which is recommended for best performance.
Q2: Do I need to wait for the resolving gel to set before adding the stacking gel?
- No. Pour the stacking gel directly onto the unset resolving gel, within 2 min, with no sealing layer. The whole gel sets within 10 min at room temperature (25°C).
Q3: My gels set too quickly when casting several at once. What should I do?
- Gelation speed depends on the amount of APS and the temperature. Reduce the amount of APS slightly when preparing multiple gels at once.
Q4: What molecular-weight range do the gels resolve?
- The 4%–20% gradient separates proteins from 10 to 250 kDa.
