A predicted band of 45 kDa appeared in the position of 95 kDa on the western blot . Shift in the position is a very general phenomena for the group of proteins it belongs to. Could you guys help me to explain it? What kind of post-translational modification can be guessed? Please help me.
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#2
bob1
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Posted 16 October 2012 - 11:33 PM
Homework?
Denaturing gel or native?
Antibody conditions?
Denaturing gel or native?
Antibody conditions?
#3
chimera
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Neutral
Neutral
Posted 17 October 2012 - 01:29 AM
It was on SDS Page. The primary antibody used in this experiment was peptide purified antibody and was used in 1:1000 dilution in 1XPBST (.1% Tween 20). The blot was developed with ECL. The experiment is reproducible.
#4
bob1
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Posted 17 October 2012 - 12:39 PM
It's a big big shift; is it a discrete band or a smear?
Are you sure it isn't a cross-reacting band from the antibody (which is a very common phenomenon with antibodies, no matter how they are purified)?
Can you see the un-modified form in the same lane? Do you have a way of reliably making the higher band appear (and what is it)?
Are you sure it isn't a cross-reacting band from the antibody (which is a very common phenomenon with antibodies, no matter how they are purified)?
Can you see the un-modified form in the same lane? Do you have a way of reliably making the higher band appear (and what is it)?
