4 replies to this topic

[zhuangyanyan]

Dear all,

I am so screwed up this morning. I am so worry right now.

I accidentally used transfer buffer as running buffer to run my gel this morning, and I found out and changed it back to running buffer in the middle, any of you has any idea what will happen? will it screw my gel????

My running buffer includes Glycine, Tris, and SDS, I took 80 ml and added 200 methanol, and 720 double distilled water

My transfer buffer includes Glycine and Tris only

Please let me know who do you think and if any of you has been through this before.

Hope to hear from you soon.

[bob1]

Your samples will not migrate very well as the SDS is the ingredient that allows separation of the proteins by size rather than charge.  The methanol will also fix the proteins to some extent, so the migration will be retarded.  You should repeat the gel and use the correct solution for running.

[zhuangyanyan]

yes, i just got the result and wanna tell everyone here. you are right. my protein does not migrate very well. i cannot see clear bands on it and my molecular marker only has 9 bands instead of 10 bands,
I donot know about the SDS part, since I found some people said that SDS has no other uses expect making your gel running linear. And there is no difference between with or without SDS.

Here is what I got. I wrote it down in case others who might run into the same situation as me

[mdfenko]

zhuangyanyan, on 18 January 2012 - 01:56 PM, said:

I donot know about the SDS part, since I found some people said that SDS has no other uses expect making your gel running linear. And there is no difference between with or without SDS.

some people don't know what they are talking about

[mdfenko]

here is a handbook on protein electrophoresis that is distributed by ge healthcare. you may find it useful and informative.

Attached Files

•  Protein Electrophoresis.pdf   1MB   124 downloads