Posted 06 December 2004 - 11:43 AM
Posted 18 January 2005 - 02:45 PM
Maybe there is a mismatch.
Edited by george@CASE, 22 January 2005 - 09:07 PM.
Posted 21 January 2005 - 11:04 AM
Posted 22 January 2005 - 09:09 PM
Here's something I found somewhere else which reiterates the above suggestion:
EASY answer as this is the most common problem with ECL type reagents. just decrease your secondary Ab concentration (the one with the enzyme). You are simply using way too much. Look at the instructions again. I know that the Sigma substrate recommends something like 1:200,000 dilution of their enzyme-ab conjugates. They mean it! I think Pierce recommends 1:20,000 and that may be a little too "rich". Amersham is off their rocker, they used to recommend 1:2,000 which was OK for their OLD substrate and poor enzyme-antibody conjugates. So read the instructions that came with the substrate and follow them carefully no matter who you purchased from. If you made your own substrate, just dilute your enz-ab conjugate about 10 fold more and see what happens.