Protein identification from bands with same molecular weight but different intensity

1D, 2D, HPLC, SDS PAGE, etc. Talk about it here.
Fazilah
Proton Member
Proton Member
Posts: 2
Joined: Mon Oct 20, 2014 8:27 pm

Protein identification from bands with same molecular weight but different intensity

Postby Fazilah » Mon Oct 20, 2014 10:18 pm

Hi All,
This is probably a very basic question, but I want to confirm if protein bands (from 1D gel) with similar molecular weight but
different intensity will end up to be the same set of proteins after LC/MS analysis? or is it possible to be different?

Thank you!

Infinity
E. Coli Lysate Member
E. Coli Lysate Member
Posts: 107
Joined: Wed Dec 21, 2011 8:22 pm

Postby Infinity » Wed Oct 22, 2014 7:21 am

It is not clear. What do you mean by saying "the same set of proteins"???

Fazilah
Proton Member
Proton Member
Posts: 2
Joined: Mon Oct 20, 2014 8:27 pm

Postby Fazilah » Wed Oct 22, 2014 5:17 pm

Hi Infinity,
Sorry if i don't explained clearly.
I plan to cut bands from a gel and identify the protein using LCMS.
If i have one band (let say sample 1) with a specific molecular weight, but low intensity, and another band (sample 2) with similar intensity but more intense, will I end up with the same proteins from these two samples? or should expect more proteins from more intense band?

Thank you!

Biomarker
Albumin Member
Posts: 88
Joined: Thu Aug 09, 2012 3:28 am

Postby Biomarker » Thu Oct 23, 2014 2:29 am

In principal yes more protein IDs with intense band. But they may be same set or different proteins..

Infinity
E. Coli Lysate Member
E. Coli Lysate Member
Posts: 107
Joined: Wed Dec 21, 2011 8:22 pm

Postby Infinity » Thu Oct 23, 2014 4:42 am

Nope, It doesn't mean anything. For example you might have very intense band but 99% of this intensity will be attributed to some high-abundant protein (albumin, keratin, etc.) - so you end up having many peptides for one protein. In an opposite way you might see almost nothing on a gel because many low-abundant proteins with similar mass and you end up having much more protein IDs.


Return to “Electrophoresis”

Who is online

Users browsing this forum: No registered users and 1 guest