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Western Blot Forum Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


Blotting large protein

Blotting large protein - Western Blot Forum

Blotting large protein - Discuss western blotting and immunoblot in the western blot forum discussion board. Ask Questions about transfers, blocking, membrane antibody incubation and exposures.


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Old 02-18-2010, 10:59 AM
Pipette Filler
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Default Blotting large protein



Hai.. I'm totally new in western blot. Is it possible to blot 230 kDa protein? If yes, can you suggest any adjustments compared to the common procedure?

Thanks a lot.
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Old 06-29-2010, 04:22 PM
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Default Re: Blotting large protein

yes, it is possible I myself am doing a 270kDa protien.
Since this is the only protien that I have worked with in westerns so far... I will just give you the protocol that I use and you can make the comparison.
First, I use 7% Tris Acetate gels along with the respective running and transfer buffer(all come from Invitrogen ). These gels only require a 1hr run @150V. Next, the transfer is done for 2 HR @ 250mA with a PVDF membrane. I usually put an ice pack in box to keep it cool, but you can also do in cold room. Then, I do the regular washes, blocking, and antibody incubations specific to my protien.
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