We want to observe the monomer and oligomer form of alpha-synuclein through the native page. But during the time of literature search we observed most of the article went through make monomer and oligomer then they did blotting but we want above both form of alpha-synuclein in tissue. Which we have done in animal model now we want to see in tissue both of the form. Please send me some protocol which is done with tissue.
You can try sds gel instead of native gel....might be you will get both on same gel.....
run the sample as a reduced and a non-reduced sample in a SDS-PAGE.
Hi Sanyan,
I will suggest you run your sample on Tris-Tricine SDS-PAGE(Gradient gel) made of 16.5%T, 6%C of separating,10%T,3%C of spacer and 4%T,3%C of stacking gel.Ensure that you run the gel with anode buffer and cathode buffer.Run at 30V for 10min until the sample goes through the stacking gel, further change to 100V. Native-PAGE is not a good choice for alpha synuclein subunits and because of its MW.
I hope this suggestion will help you.
All the best!
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