QuickRun™ Running Buffer – Protocol
QuickRun™ running buffer is a proprietary formulation specifically developed to drastically increase the separation speed without loss of resolution typically observed after hour long migrations. This buffer is compatible with all protein gel chemistries where the pH of the separator gel is = 8.0 and is compatible with all downstream Western blotting applications.
QuickRun™ buffer is formulated to extend the separation range of a linear gel so casting gradient gels is not needed anymore! The separation ranges will be as follow:
- An 8% linear gel will behave as a 4-12% gradient gel;
- A 10% linear gel will behave as a 8-16% gradient gel;
- A 12% linear gel will behave as a 4-20% gradient gel;
- A 14% linear gel will behave as a 10-20% gradient gel.
The QuickRun™ formulation is very robust and tolerant of a wide range of, otherwise, unacceptable electrophoresis products and solutions. Contaminated distilled water, bad quality acrylamide, expired precast gels and gel solutions can all be used if QuickRun™ is the running buffer. There are limits to what QuickRun™ can work with in these instances (exclusions include: physical damage to a gel, bubbles or cracks in the gel matrix, dehydrated gels).
Prepare a 1X running buffer solution by diluting the stock buffer with distilled water
- Fill your electrophoresis chamber. Note that if you want to run the gels at higher voltage (higher than 200 V), make sure to completely fill both electrophoresis chambers to the maximum level allowed so that there is adequate heat transfer and even gel migration.
- Install your gel(s), flush the wells with running buffer and load your samples
- We recommend, when you use this buffer for the first time, calibrate your gels with QuickRun™ and a prestained MW marker to see the migration length and stop it as you need
- Run under your preferred conditions. Please be careful, the migration is faster than your regular buffer
As a guide, we recommend the following conditions*
175 V, for about 25 min OR 220 V, for about 20 min
Maximum recommended voltage 250 V
(to prevent over-heating of the gel)
This buffer is re-usable, but be careful, if you migrate long and have proteins going out of your gel, they can be carried over to the gels run with the used buffer.
*Note: The migration time may vary with the concentration of your gel, cassette size (the above recommendations are for an 8 cm gel), your equipment, ambient room temperature and the ionic strength of your sample buffer.