Hello,
I obtain western blot bands at desired region on the membrane but there is no space between the bands (attaching image for reference), may i know what could be the reason?
TIA
Do you know how much protein you added? The bands also look pretty intense so maybe you overloaded the gel?
The bands on the gel are not continuous. You still see the boundaries between the 6 samples, they have just migrated closely together. This is a normal effect of SDS-PAGE, since the differences in salt concentartions and pH between the gel and the samples lead to a broadening of the samples (electric current pulling the proteins not only downwards, but also sidewards at the edges), which is usually stooped when the sample from the neighboring well interferes with sidewards movement. If you need to have free space beween the samples, leave one lane free (or better fill it with empty loading buffer) between the samples.
Sebastian Fiamma These are extracted histones from the cells and loaded 5 microgram/lane
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