Why is BN-PAGE conductivity lower than expected?

If you are experiencing lower conductivity than expected during your BN-PAGE run, there are a few potential reasons that could contribute to this issue:

Buffer Preparation: Ensure that the upper buffer and lower buffer used in the gel system were prepared correctly, following the established recipes. Double-check the concentrations of the necessary components, such as Tris and glycine, and make sure they were accurately measured.

Buffer pH: Confirm that the pH of the buffers is within the appropriate range. Deviations from the optimal pH can affect the conductivity of the buffers and subsequently impact the efficiency of the electrophoresis run.

Salt Contamination: Check for any salt contamination in the buffers or electrode chambers. Salt contamination can occur if there was insufficient rinsing of the chambers or if there was cross-contamination from previous experiments. Thoroughly clean the chambers and ensure proper rinsing to eliminate any residual salts.

Gel Quality: Examine the gel itself to ensure its integrity. Look for any cracks, air bubbles, or irregularities in the gel matrix that could affect the conductivity. Additionally, verify that the gel is securely positioned within the gel chamber and that it is making proper contact with the electrode buffers.

Electrode Placement: Confirm that the electrodes are correctly positioned in the gel chamber. The positive electrode (anode) should be placed near the sample wells, while the negative electrode (cathode) should be positioned at the opposite end. Incorrect electrode placement can result in poor conductivity and incomplete gel running.

Age of Reagents: Check the age and quality of the reagents used in buffer preparation. Expired or degraded reagents can affect the conductivity and performance of the buffers. Ensure that all reagents are within their expiration dates and stored properly.

By carefully investigating these potential factors, you should be able to identify the underlying cause of the low conductivity during your BN-PAGE run. Addressing the specific issue will help you resolve the "Error no load" message and improve the efficiency of your gel electrophoresis.

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Debjyoti Boral

I am facing the same issues while using the tricine imidazole buffer system for BNPAGE. I hope you resolved the issue.

I want to use the Tris-glycine buffer system.

Could you by any chance share the composition of the buffer systems used as Cathode and Anode buffers in your successful experiments?

Also did you face a smear issue for the samples run?

Any responses are welcome.

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