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NOS western blot problem

NOS western blot problem - Protein Forum

NOS western blot problem - Protein Forum

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Old 02-04-2007, 01:43 AM
Wang, Hongyan
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Default NOS western blot problem

Hi, Anna,

I found that I now have the same problem as you had, except the gene
that do well is SOD1, actin as housekeeping gene. Could you please tell
me what happened to your samples and how did you solve it. I would be
grateful for suggestions. Thanks.


Anna Durrans anna.durrans at physiol.ox.ac.uk
<mailtoroteins%40iubio.bio.indiana.edu?Subject=N OS%20western%20blot%20
Fri Jul 13 08:39:40 EST 2001

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Hi everyone,

I have been running westerns for eNOS, nNOS and iNOS successfully using
various rodent organ tissues. I use actin as a housekeeper.

In order to show reproducibility I recently re-ran the exact same
again, along with some freshly made lysates. Strangely I found that the
actin signal was very much weaker in some (not all) of the original
as wells as in the fresh lysates.

I re-ran all these again, and while there is a pretty stong actin
is still weaker in those tissues which before showed a weak actin

I am not sure what could have happened. The NOS signal in ALL the
was fine - so it is not the lysates themselves but possibly the actin
specifically which has degraded.

Has anyone else had this problem? Does anyone know what could have

I would be really grateful for any suggestions.


University Laboratory Department of Biochemistry
of Physiology South Parks Road
Parks Road Oxford OX1 3QU
Oxford OX1 3PT Tel: 01865 275275/275273

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