Dear ResearchGate community,

I am currently facing an issue with the solidification of my SDS-PAGE electrophoresis gels. Despite following the standard protocol, the gels fail to solidify properly, which is hindering my protein separation experiments. I would greatly appreciate any insights or suggestions to resolve this problem.

Here are the details of my gel preparation protocol:

I prepare the gel solution by mixing the following components:

Acrylamide

SDS

TEMED (N,N,N',N'-Tetramethylethylenediamine)

Ammonium persulfate (APS)

After thoroughly mixing the components, I pour the gel solution into the casting tray and insert the comb to create wells for sample loading.

I allow the gel to polymerize for the recommended time, usually 30-60 minutes, at room temperature.

However, despite following these steps meticulously, the gels do not solidify completely and exhibit a semi-solid or liquid consistency. As a result, I cannot load my protein samples for electrophoresis.

I have considered the following factors that might contribute to this issue:

Aging of the acrylamide and bis-acrylamide: I ensure that my acrylamide and bis-acrylamide solutions are freshly prepared. I have also checked their expiration dates to ensure they are not expired.

Incorrect ratio of reagents: I carefully measure the quantities of acrylamide, SDS, TEMED, and APS according to the protocol. However, it is possible that I have made an error in the measurement.

Incomplete mixing of components: I thoroughly mix the components to ensure proper dispersion, but there is a possibility of inadequate mixing leading to incomplete polymerization.

Temperature variations: I perform gel polymerization at room temperature, which is typically around 20-25°C. However, ambient temperature fluctuations may affect the gel solidification process.

Considering these factors, I am seeking advice or suggestions on how to troubleshoot this problem and achieve proper gel solidification for my SDS-PAGE experiments. Any insights or recommendations would be highly appreciated.

Thank you in advance for your time and assistance.

Sincerely,

Sachinkumar

helix chloro pvt ltd