Western blot bands look like 'upside down smiles' - (Mar/13/2006 )

Hi,

Please help!
Recently my Western blots bands have turned into upside down smiles. Previously they have always run ok. Does anyone know why this could be? Maybe contamination of buffers? I've tried making up fresh buffers but it hasn't made any difference. Perhaps the solutions we use to pH buffers are contaminated? Please let me know if you have seen this before and what you did to correct it.

Many thanks

Becky

i think it's probably aproblem of polymerization. Preparing fresh APS will maybe do the job.

hi,
Try using fresh APS and TEMED. next suggestion is while polymerization keep the setting in 37C for one hour. it improves the polymerization process.

gud luck

Hi smiling band has two possible reasons:
1) protein overload
2) too many salts in the sample - I homogenize in 30 mM TrisHCl and have perfectly straight lines,
I hope this help

get some ethanol, and a kim wipe and *VERY* carefully clean the metal wire at the bottom of the apparatus you are running within.

Smiles and frowns tend to be caused by other stuff, (I have seen them from big samples in small wells, and from old gel) but dirt on that wire can cause shapes other than straight.