i am having trouble with my sandwich elisa with streptavidin hrp, 1 step tmb substrate.
6/10 plates i have run have presented the same issue and I have been observed by a collegue who confirmed its not my trchnique causing the odd results.
So, my plate develops perfect except for rows A,B, and C down the entire plate including std curve (500ng). its not a plate washer issue as we rotate our plates 180 and perform a second wash each step to overcome washer variability. all my incubations are at 37 for one hour. originally i thought the issue was stemming from stacked plates but I stopped that habit and results were the same.
so again, everything to the right of row B/C develops perfect but everything to the left doesnt develop at all. any ideas?
Edited by matty2431, 19 August 2009 - 12:16 PM.