I want to see a 16kDa band of my protein in SDS PAGE. Whenever i run the gel, the band in the upper part of resolving gel were very sharp and clear however the lower part where i supposed to get my 16kDa band was very hazy and not resolved properly. Kindly suggest what to do in this case. I am running my gel at 25mA in 18 x 16 cm glass plate
Hi,
I agree with Shiv Shankar. It is recommended to use a gel with a concentration of 10% or 12% and a voltage of 120-200 V. If the voltage is too low, small molecular weight protein bands will diffuse.
Hope it helps!
I generally use 15% of resolving gel and 5% of stacking gel. The bands in the upper portion resolved properly however in the lower portion marker (protein ladder) itself gets distorted bands. I don't know why its happening like it?@Shiv shankar and @ Roger Davis
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