Hi,
I run SOD zymogram by using 4-20% native gel and with native sample buffer. I lysed my cells in Tris-Cl buffer. I homogenized cells with a homogenizer and then spin them down to extract protein. Then I load them not boil them (I don't know it is a problem not to boil but I run without boiling). I did incubate them with 0.01% NBT then washed with Di water then incubate them with riboflavin solution again was with Diwater then I expose it to fluorescence light which is blue and I have the result in the attachment. I don't have achromatic band. I have a purple band. Where do you think the problem is? Do you think blue flourescent loght is a problem or the solutions I used is a problem? Also purple band size is not correct we are supposed to see it at orange band but we see them around 10kD which is blue line.
Thanks fr your help,
Tugba
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