I am working on a recombinant protein using SDS-PAGE. I used Coomassie brilliant blue for visualization of the respected protein bands. The voltage was set at 150V and the gel was run for 1 hour. Acetic acid was used for decolorization of the gel.
All SDS-PAGE buffers were appropriately prepared and used fresh. However, there is a weird curved line at the center of the gel. How can I get rid of it?
Your samples are not resolved well. Try running your sample on a higher percentage of acrylamide at a constant current- 20mA for one gel. Use fresh buffer and make sure that the electrode assembly is not leaking. Destain the gel on a shaker. When destaining two gels make sure you've given enough destatining solution and they aren't stuck together.
Since the line is not limited to the lanes, but is also seen between them, it probably came from a contaminant in the running buffer, or in the buffer reservoir. The most common contaminant is keratin, which comes from skin and hair, but it could be some other contaminant. Make fresh running buffer. Be sure to use thoroughly clean vessels, rinsed with distilled water to remove any residues, and wash out the PAGE apparatus thoroughly with distilled water.
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