FlexiRun™ – Protocol
FlexiRun™ is a proprietary formulated premix of acrylamide gel for easy, fast (10 Min.), safe, and high resolution performing gel. The proprietary premix delivers crisp sharp and non-smiling bands. Polymerization speed adjustable to your needs.
FlexiRun™ delivers below unique benefits:
- Easily cast ANY % gel in 10 Min. by diluting in water only.
- Compatible with ANY buffer system including QuickRun™, Tris Glycine (with or without SDS), Tris Tricine, Tris Taurine, Tris MES, Tris HEPES, or Tris MOPS. For best gradient-like separation and fast 15 Min. protein gel run, use together with QuickRun™ Running Buffer (P/N: ID1591).
- Sharp bands with any sample type including direct loading of crude lysates.
- Adding extra SDS to the gel solution is not needed with this formulation.
This protocol is written for mini 10 X 10 cm or 8 X 8 cm gel cassettes, but can be adjusted for any size cassette, just by increasing the volumes accordingly. It is easy to pour two mini gels at the same time.
For each mini gel – prepare a 10 ml of separator gel solution
- Determine the percentage of gel you want, and divide the value by two
- The value you obtain is the amount of FlexiRun™ required
- Add sufficient water to give 10 ml final volume
- Add 10 µl TEMED
For every one or two mini gels – prepare a 10 ml of stacking gel solution
- A stacking gel concentration can be between 4 to 6%, we recommend 5%*
- Take 2.5 ml of FlexiRun™
- Add 7.5 ml water for a 10 ml total volume
- If you want to easily see the well separators, add 10 µl of Blue acrylamide dye (cat ID2001) to the stacking
- Add 10 µl TEMED
Pour the gel(s)
- Add 100 µl** of fresh 10% APS in both the separator and stacking solutions
- Mix slightly by one inversion of the tube
- Pour the separator in the cassette up to the level you want (planning for a 1 cm stack)
- Using a 1000 µl Pipette, immediately add, drop by drop covering left to right of the separator gel surface, the stacking gel solution to fill the cassette
- Place the comb in position and allow polymerization for 5 to 10 minutes
* Polymerization of a low concentration gel can leave the well separators weak for a while, and they can move when removing the comb. It is easy to put them back in place with a needle, or increase the stacking concentration to 5.5% or 6% or increase the polymerization time
** This protocol has been calibrated for RT defined as 20˚-24˚C, at higher temperatures, the polymerization will be faster. Following this protocol, you have about 2 minutes to pour the gels. This can be difficult if you want to pour several gels simultaneously. However, it is easy to slow the polymerization time: simply reduce the amount of APS 10% to 75 µl, to get about 5 minutes before polymerization. Reduce APS 10% to 50 µl for about 10 minutes of polymerization. Be aware that the stacking gel solution is slower to polymerize than the separator gel solution.