Recently I run a Glycine-SDS-PAGE to detect the autophagic protein LC3-I and II (16 and 18Kd) . I loaded the recommended quantity of sample: 40ug. The transfer was OK, but my bands appears diffuse with a long shadow. I run to 80V until the limit between gels, and then to 120V, 3hrs in total. I prepared new and fresh buffers to make the gels, but nothing..I get the same result.
Which other aspect-change-mistake-trick should I consider?
If you need to see my recent Western Blot to estimate what might happen (like a doctor), I have attached the image in this message.
Thanks in advance.
uggly, non-focus and diffuse bands in my Western..
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