white bands in wb - (Dec/11/2006 )
i would appreciate ur help in this
in today's WB detection i found white bands in a very high background even in 1 seond exposure of the film?????
i didnt change any thing at all as yesterday for example....
i didnt change the first or second antibody or its concentartion or anything at all????
my samples are the same as before...
first time really to face this problem, i tried to wait 15 minutes after the first exposure and try again but my film was nothing there at all even the background.
i am using ECL detection .
i appreciate ur help.
This happened to me when once I switched from ECL to ECL advance (which is even more sensitive than ECL plus) without diluting enough my HRPsecondary. I got big white spots with less than 5 seconds exposure
So probably your white bands are also due to too much secondary, but if you didn´t change anything... it´s strange... Perhaps you took a wrong ECL (in case of somebody in the lab uses a more sensitive one and you confused with yours )
Let´s see what the others said.
In case it helps a bit, here it´s discussed this problem: http://www.protocol-online.org/biology-for...osts/11707.html
i know what u said and i did search the net for answers but the strange thing is that i didnt change anything from my usual and i was just repeating this western as i didnt include control in the last one ....
we are only 2 in the lab that use this ECL and its only one kit.
thankx for ur reply, waiting with u.
Could it be that your signal is now too high because you included your positive control whereas before you had your protocol titered to a lesser signal? Just guessing!