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Guy,
I slightly disagree with you point of view and I second Martin's opinion.
If you e.g. go to the Millipore website (http://www.millipore.com; no commercial interests) and you search for tubulin antibodies you fill find a variety of products. For each of them you find a key application like e.g. Immunocytochemistry, Immunohistochemistry, Western Blotting or Flow Cytomertry. Some of them are optimized/tested for only one application others can be used for all of them. So there seems to be a difference and it is worth while checking before.
Best regards
Arne
> -----Original Message-----
> From: Confocal Microscopy List
> [mailto:[log in to unmask]] On Behalf Of Guy Cox
> Sent: vendredi 8 mars 2013 17:02
> To: [log in to unmask]
> Subject: Re: antibodies for immunofluorescence
>
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>
> Martin,
>
> I must say I am very worried by your statement "Antibodies that work
> for one application (e.g., westerns) may be completely unusable for another
> (e.g., IF)". Can you give me a biological reason for this? It would seem to
> make most cell biological work impossible. I would assume that most of us
> work the same way as me - ie using microscopy to identify the location, and
> chromatography to characterise the protein. If you don't use the same
> antibody both times you will just be generating nonsense!
>
> Guy
>
> ________________________________________
> From: Confocal Microscopy List [[log in to unmask]]
> on behalf of Martin Wessendorf [[log in to unmask]]
> Sent: 09 March 2013 02:31
> To: [log in to unmask]
> Subject: Re: antibodies for immunofluorescence
>
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>
> Dear Michal--
>
> On 3/8/2013 2:36 AM, Micha³ Majkowski wrote:
>
> > I do not know if this is a good place to address my problem as it
> > regards immunofluorescence. The problem is: we have in the lab
> > antibodies dedicated to Western Blot (according to manufacturer; Abcam).
> > We used them to IF and we obtained some nice images. What do you think
> > about such data? Should it be confirmed by IgG dedicated to IF? Thank
> > you for help.
>
> The quick and safe answer is that you can never trust any antibody for any
> purpose without first characterizing it thoroughly.
>
> Antibodies that work for one application (e.g., westerns) may be completely
> unusable for another (e.g., IF)...or they may work perfectly.
> A starting point is to compare the labeling that you obtain, to the labeling
> that's previously been published. The "gold standard" is to demonstrate that
> the labeling is not observed in a knockout animal.
>
> Cliff Saper at Journal of Comparative Neurology published an editorial on
> antibody use that may be helpful. It can be found at:
> http://www.wiley.com/legacy/wileyblackwell/images/antibody_editorial .
>
> Good luck!
>
> Martin Wessendorf
>
> --
> Martin Wessendorf, Ph.D. office: (612) 626-0145
> Assoc Prof, Dept Neuroscience lab: (612) 624-2991
> University of Minnesota Preferred FAX: (612) 624-8118
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