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Author Topic: Oligodendrocytes and Microglia  (Read 10654 times)

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Offline badchad

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Oligodendrocytes and Microglia
« on: August 03, 2005, 04:45:28 PM »
Wish I had found these forums about 6 months ago before!

I have a question about the selection of two different antibodies.  I'm using male F-344 rats and paraformaldehyde fixed, coronal sections about 40 microns thick.

1. I'm looking for a specific oligodendrocyte marker.  I've pretty much narrowed my selection to either an O1 antibody, or a CNPase anitbody.  Both are supposedly selective for mature oligodendrocytes, I would be interested to hear anyone's opinions/experiences with either antibody.

2. I'm also looking to label microglia.  This should be easier than the first one.

What I'm looking at is the expression of the c-fos protein in rat brain in response to novel stimuli.  While initially I was using c-fos as a marker of neuronal firing, I found that the expression didn't occur in neurons (from doubel-labeling with a neurofilament and c-fos antibody).  Morphologically, the cells appear to be oligo's but I'm trying to confirm this.  

I'll be using some AlexaFlor antibodies as secondaries.  ANy and all help is appreciated.

Oligodendrocytes and Microglia
« on: August 03, 2005, 04:45:28 PM »

Offline richard03

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Oligodendrocytes and Microglia
« Reply #1 on: August 03, 2005, 11:38:57 PM »
I have used CNPase from Chemicon based the following protocol with good success:

http://www.ihcworld.com/_protocols/antibody_protocols/cnpase_chemicon.htm

For microglia, I have used Lectin RCA-1 from Vector. It label microglia and blood vessels. But it should be very easy to distinguish microglia from blood vessels. Here is the protocol I used:

http://www.ihcworld.com/_protocols/special_stains/microglia.htm

You may also try antibody staining such as Iba1 for microglia and this should be more specific. Here is Iba1 staining protocol for microglia:

http://www.ihcworld.com/_protocols/antibody_protocols/iba1_santa_cruz.htm

http://www.ihcworld.com/_protocols/antibody_protocols/iba1_wako.htm

Hope this helps.

Richard

Offline richard03

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Oligodendrocytes and Microglia
« Reply #2 on: August 03, 2005, 11:41:22 PM »
By the way, if you want to identify neurons, use NeuN - a neuron specific marker and the protocol is here:

http://www.ihcworld.com/_protocols/antibody_protocols/neun_chemicon.htm

Offline Peter Canoll

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Oligodendrocytes and Microglia
« Reply #3 on: September 25, 2005, 07:09:41 AM »
Ox42 works well for microglia.
olig2 will label the nuclei of cells of the oligodendrocyte lineage, progenitors and mature oligos
Other antibodies that we have used to label oligodendrocytes CC1, MBP, O1, CNPase, of these CC1 is nice because it labels the cell bodies and makes it easy to localize and count.
 am looking for a rat-specific nuclear antigen for IHC

Oligodendrocytes and Microglia
« Reply #3 on: September 25, 2005, 07:09:41 AM »