Western not working
Posted 03 July 2013 - 09:12 AM
We've got some serious and weird problem with our western blots. All our western blots have stopped working. It's not the gel or transfer as we get good staining on the membrane by ponceau. It's also happening for all our primary antibodies, not one, so it's unlikely to be that. We've got a new secondary HRP and new ECL reagent. The westerns give no signal on film - then give a good signal and then give no signal - all with no warning or any reason I can think of. Has anyone got any ideas?
Posted 03 July 2013 - 09:36 AM
Hope this helps... is a very quick test and fun
edit: Also (though less fun), check the pH of your wash and incubation buffers as this might affect the antibodies, and I assume this is common for all your westerns not working.
Edited by almost a doctor, 03 July 2013 - 09:37 AM.
Posted 03 July 2013 - 12:22 PM
Posted 04 July 2013 - 02:01 AM
Posted 04 July 2013 - 08:49 AM
talent does what it can
genius does what it must
i used to do what i got paid to do