Dear Tuba Öz

There can be several possibilities for frowning bands.

• Use fresh running buffer. Fill the tank as indicated. If you still get frowning bands, cover the whole tank with 1X buffer.
• The voltage might be two high for the type of your gel. Lower the voltage.
• Run the stacking gel in lower voltage for half an hour approximately, then increase the voltage when the bands come to resolving gel.
• Make a homogeneous mixture of the gel before casting.
• If the running buffer gets warmer during running the gel, use ice to cover the tank or run it in a cold room.

Best wishes.

Dear Tuba Öz

Try using precast gel if you have access to one. Also, how did you prepare your running buffer?. You can as well reduce the voltage 100v when your sample is still in stacking Gel.

Dear Satyendra Mondal

thank you for answering.

I can use fresh 1x tris-glycine sds running buffer. I fill the tank as indicated. Then I run the electrophoresis at 100V, when my sample is still in stacking gel and then I run the electrophoresis at 124V for resolving gel.

Dear Abdullahi Hassan Ndanusa

thank you for answering. I used fresh 1x tris-glycine sds running buffer. Then I run the electrophoresis at 100V when my sample is still in stacking Gel. And then I run the electrophoresis at 124v for resolving gel.

Frowning bands in SDS-PAGE can be caused by several factors, including uneven sample loading, overloading of the gel, running the gel for too long, and uneven distribution of the current during electrophoresis. Here are some possible causes and solutions:

By troubleshooting these possible causes, you may be able to improve the quality of your SDS-PAGE results and eliminate frowning bands.

These video playlists might be helpful to you:

https://www.youtube.com/playlist?list=PLzBiAPKi8vOPbE7Db9mKwloe34E8rXXEU

https://www.youtube.com/playlist?list=PLzBiAPKi8vONSqjcweFWHMRy13y3484bE

https://www.youtube.com/playlist?list=PLzBiAPKi8vOMheKwb0mu1dcZRaeGUFSlw

Nikolay Klyashtornyy Thank you, I will go through each steps. these video playlists are wonderful.