Very odd western bands - (Aug/09/2007 )

Hi everybody,
I was hoping to get your learned opinions on some western issues a person in our lab is having. Ive done a number of westerns but have never run into or seen this problem before. Our running buffer is commercial and have had no problem with it previously- the sample buffer we use is Laemmli buffer and others in the lab use the same stock with no such issues. I was thinking it might be something to do with the protein but am at a loss. I have attached a pic of the kind of western produced.
Thanks for any help!

Caroline

Just wondering...

Does this person use commercial SDS-PAGE gel?

If yes, the gel may be expired.

If not (he/she made the gel),
then the gel looks like it is split into two portions...with different % of acrylamaide.

You may wish to ask that person how he/she make a SDS-PAGE gel.

Hope this may help.

Hi,
Thanks for the rapid reply!
The membrane was cut in order to probe two seperate antibodies- I just forgot to remove the top section before posting.
We do buy commercial gels and they were purchased only about two weeks ago. Company claims they have a shelf life (4oC) of 1 year.

Thanks
Caroline

Hi,

has the sample a lot of salt or DNA?
These compounds can cause band distortion on sds-pages...
Try an dialysis to remove salt or melt the sample before appling it on gel...

Sorry for the bad English (I´m still learning...)...
Best wishes, from Brazil!

I also think that this phenomenon is due to components in the sample, because the marker bands are nice.