Introduction:

Blue Native PAGE Running buffer is formulated for separation of proteins or protein complexes while retaining their native structure through gel electrophoresis on blue native gels. BN-PAGE substitutes Coomassie brilliant blue G-250 for SDS to make the protein complexes negatively charged and separated in the gel according to the molecular weight of each complex. This product is a 5× concentrated solution, which needs to be diluted to 1× working concentration before use.

Storage: Store at 4℃, valid for 1 year.

Protocol (for reference only):

1. Measure 100 mL of 5× anode running buffer, pour it into a clean beaker, add deionized water to about 400 mL, and mix well.
2. Make up to 500 mL in the graduated cylinder with deionized water, which is 1× working anode running buffer and can be used after mixing.
3. Prepare 1× cathode buffer A and 1× cathode buffer B in the same way as the anode buffer.
4. Load your protein samples into the wells of BN-PAGE gel before adding running buffer because the buffer obscures visibility and may interfere with accurate loading.
5. Add 1× Blue Native PAGE Cathode Buffer A to the inner buffer tank to cover the wells.
6. Add 1×Blue Native PAGE Anode Buffer to the outer buffer tank of the electrophoresis device, assemble, and connect to the inner buffer tank.
7. Run the gel. When the sample runs to 1/3 of the gel, replace the 1× Blue Native PAGE Cathode Buffer A with 1× Blue Native PAGE Cathode Buffer B, and continue the subsequent electrophoresis.

Notes:

1. 1× working buffer should not be used for more than 2 weeks after preparation.
2. This running buffer is suitable for Blue Native PAGE gels, and should not be used for gels with other chemistries.
3. For research use only. Not for use in diagnostic procedures.
4. For your safety and health, please wear personal protective equipment and clothing when operating.