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Separating proteing with difference of 5Kd on western blot

Posted: Mon Dec 17, 2012 8:14 pm
by Preeti
Hello everyone,
I am kind of stuck here with two proteins with almost similar molecular weight and I want to do Western blot of them as it is a
requirement of the project. Can someone guide me?? the weight is 90kd and 95 kd.
Thanks,
Preeti

Hi

Posted: Tue Dec 18, 2012 12:02 am
by Biomarker
Hi Preeti,

What percentage of SDS-PAGE you are using? Try to alter the percentage of acrylamide. I hope this will help to you..

Biomarker

Preeti wrote:Hello everyone,
I am kind of stuck here with two proteins with almost similar molecular weight and I want to do Western blot of them as it is a
requirement of the project. Can someone guide me?? the weight is 90kd and 95 kd.
Thanks,
Preeti

Posted: Tue Dec 18, 2012 3:03 pm
by Preeti
Hello Biomarker,
I am using 4-17% gradient gel as i have four more proteins which I need to study on same blot. All to gather i am studying six proteins and one as control common housekeeping protein.
If you have any idea to sort this problem please do snuggest me. Thanks for your reply.
:) Preeti

Posted: Tue Dec 18, 2012 6:50 pm
by Sidney
Hi, Preeti,

I would like to suggest you perform western blotting from two SDS-pages, because antibody has very high specificity to assure the right protein being annalyzed.

Sid

Hi

Posted: Wed Dec 19, 2012 2:23 am
by Biomarker
Hi Preeti,

Try 8-16% and 10-16%.
Good luck....

Cheers...
Biomarker

Posted: Wed Dec 19, 2012 4:24 pm
by Preeti
Thanks for your suggestion Biomarker!!! :)
Would inform you the outcome and I wish either of them will work!!!
Cheers, :D

Preeti

Posted: Wed Dec 19, 2012 5:50 pm
by Preeti
Thanks for your suggestion Biomarker!!! :)
Would inform you the outcome and I wish either of them will work!!!
Cheers, :D

Preeti