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Help needed w/ buffer stability

frandenm at TCPLINK.NREL.GOV frandenm at TCPLINK.NREL.GOV
Mon Jul 18 13:18:54 EST 1994


I also used o-dianisidine and H2O2 as substrates in western blots using
peroxidase-labelled antibodies.  I made up my solutions fresh, right before 
use -- do not store.

Through trial and error, I also discovered that glassware had to be clean.
When I used plasticware, I would get variable staining, which I assumed to
be due to the presence of inhibitors for peroxidase in less clean labware.


______________________________ Reply Separator _________________________________
Subject: Help needed w/ buffer stability
Author:  mmccrear at magnus.acs.ohio-state.edu (Marvin R McCreary) at smtp
Date:    7/18/94 10:54 AM


I am using a 0.05 M Potatssium Phosphate Buffer containing o-dianisidine and 
h2o2, as a substrate containing solution for a myeloperoxidase assay.  After 
receiving the solution 10 days ago, their was particulate matter in the 
solution, and the myeloperoxidase activity of a standard solution was decrease 
(presumably due to a bad buffer solution).  The buffer solution had been 
refrigerated as directed.
     
Does anyone have any suggestions, or experience with these buffers so that I 
could avoid this problem?
     
Thanks in advance.
     
McCreary.14 at osu.edu
     




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