Long Sds

July 8, 2008 By Leave a Comment

Long Sds
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Please help me analyze my SDS-Page! Your help is much appreciated!?

I ran an SDS-Page containing reduced and non-reduced proteins of Insect SF9 cell lysate, human IgG, beta amylase, and BSA. There is a standard size marker as well. Here is where I get confused, what causes the huge long smudgy trails of a protein? Why is my homotetramer beta amylase showing up at approximately 50 KDa for both reduced and non-reduced when my professor told me the size of beta amylase is 210 KDa? Are there different kinds of beta amlyase? I understand that denaturing means the proteins unfolds and that reducing means the disulfide bonds are broken. I don't understand how this would change the size of my protein. Also, why does my human IgG have a couple of bands instead of one in an SDS-Page.

My professor unfortunately does not understand my questions due to a slight language barrier, thus, it is a battle to ask even the simplest of questions. I really want to understand my SDS-Page.

I'm not sure, but it sounds like the first step is you need to reduce the volume of protein you are loading. If there is too much, then the proteins can't untangle well enough and they make a big mess on your gel, and make it almost unreadable.

It can slow them down and make them move to places you don't want them to.

Also, be careful you haven't tainted your sample by touching it or breathing in it or anything because then you add your own body proteins to it too.

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