H.Veldman-2 at neuro.azu.nl
Fri Feb 16 03:30:35 EST 2001
= Henk Veldman <H.Veldman-2 at neuro.azu.nl> wrote:
= > Angie schreef:
= > > Does anyone know of a robust and fairly clean antibody to use for
= > > histochemistry that localizes to the cytoplasm of glia, preferably
= > > astrocytes, both activated and non activated?
= > >
= > > Thanks for any help!
= > >
= > Almost any producer of commercial antibodies will have one or more
= > preparation in its catalog against GFAP (glial fibrillary acidic
= > protein), both polyclonal and monoclonal. Try DAKO, Sigma, Chemicon,
= > well . . . any.
= > We are using the DAKO polyclonal antibody on sections of fixed rat brain
= > and on cultured cells to identify and locate astrocytes, much to our
= > satisfaction.
= > Henk
= Yes, we are waiting on a new batch of GFAP, however it was our
= understaning that in tissue it stains primarily activated astrocytes. (Am
= I wrong about that?) We are trying to find an antibody that will stain
= both activated and non-activated astrocytes in tissue sections. We have a
= reaction product from another histochemical stain that we are trying to
= co-localize. The first reaction is staining nuclei throughout the cortex,
= and we are trying to find out if the nuclei are from neurons or
= astrocytes. We have tried S100-beta, but as it results in nuclear
= staining as well, it's a bit problematic.
The anti-GFAP antibodies should stain all astrocytes in tissue sections.
However, the intensity will vary. Those in the spinal cord stain stronger than
the ones in the brain and activated astrocytes stain stronger than others.
Usually the staining will be strong enough, depending on things like fixation,
tissue processing and detection method.
If not, you could try antibodies to vimentin, as these will visualize even the
weakly staining unactivated astrocytes, but will not react with neurons.
Laboratory for Experimental Neurology (NMZ)
University Medical Center Utrecht (AZU)
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