[Histonet] Re: beta tubulin

Carl Hobbs carl.hobbs <@t> kcl.ac.uk
Tue Aug 14 13:37:50 CDT 2007

you don't mention what fixing solution/ duration you use......
I have not used the primary Ab you mention: however, as it is not a 
neurone-specific Ab, you need to use another neurone-specific Ab to confirm 
that what you are demonstrating are neurones ( sure, morphology helps but is 
not neccessarily definitive) OR switch to a neurone-specific beta tubulin 
( beta III tubulin- neurone specific).
Otherwise , it seems to me that your technique is about right, provided that 
you are permeabilising after fixation but before primary Ab incubation. You 
will need to permeabilise to optimise staining, dependant on all the other 
variables in your technique, that you haven't mentioned
NB: Can't comment on appropriate dilution of your primary Ab but your 
Alexafluor 488 should be fine......altho' I use it at 1/1000 dilution 
Have a look at IF pics of neurone -specific beta tubulin here, for example : 
Best of luck

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