In this electrophoresis , i'm using serum samples without any treatment, mixing them with sample buffer in a 1:1 ratio. Te run is performed at 150v for 1 Hour, an this is the result after staining.Has anyone experienced a similar situation? do you have any recomendations? Thank you.
Try increasing the percentage of the gel and reducing the concentration or volume of the sample
Given that the ladder on the lhs of the gel is trying to work well and the single sample on the rhs of the gel is spreading in a similar shape to the mess in the middle it looks like some salt or other contaminant in your samples is causing the samples to spread and the central samples being close to each other the effect is much greater I would not worry about the gel and pay particular attention to what is remaining or being added to your samples prior to running them but in answer to your question,,no I have never seen this effect before
Have you quantified the amount of protein in your samples?
These oval bands that spread over neighboring lanes usually suggest massive amounts of protein loaded into the gel. Human serum is very rich in proteins, especially albumin. I wonder if this is the dominant band we are seeing in your gel.
Thank you for the response. Yes, it is an effect due to the concentration of the samples.
Такое впечатление, что анализируемые белки диффундируют в соседние лунки и даже в кюветный буфер. Попробуйте разводить сыворотку 60%-ным раствором сахарозы в кюветном буфере в соотношении: (сыворотка : р-р сахарозы = 1 : 10), и наслаивать пробу в лунки концентрирующего геля буквально 1 мм или даже меньше. Должно помочь :-)
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