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Author Topic: Need glia marker  (Read 8713 times)

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Offline caldera

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Need glia marker
« on: May 24, 2004, 10:23:50 PM »
I am looking for a marker for the glia in the dorsal root ganglia(DRG) of mouse embryo. Does anyone have good suggestion? Thanks

Need glia marker
« on: May 24, 2004, 10:23:50 PM »

Offline Tony

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Need glia marker
« Reply #1 on: June 03, 2004, 03:41:05 PM »
What about GFAP for astrocyte, Lectin RCA-1 for microglia?

Tony

Offline mhirano

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Good anti-GFAP?
« Reply #2 on: July 21, 2004, 04:07:50 PM »
I have tried to stain FFPE sections of mouse brain (cerebrum) with rabbit anti-GFAP antibody (Zymed, 18-0063) by IF. I observed fluorescent strings, but very sparsely distributed (especially compared to DAPI-stained nuclei). Consistent with manufacturer’s statements, neither HIER nor PIER improved the staining results. Since I could not see fluorescent strings in control sections stained without anti-GFAP, I believe that this antibody worked. Are glial cells so rare in mouse brain? Or, are most nucleated cells neurons in cerebrum?

Anyway, I should like to try another anti-GFAP antibody or RCA-1. If you know a good anti-GFAP antibody or commercially available RCA-1 with “tag” (e.g. biotin), let me know, please.

Offline Jossy

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Need glia marker
« Reply #3 on: July 22, 2004, 08:05:35 PM »
Hi, I have used anti-GFAP from Dako (rabbit policlonal) and works very good in DAB IHC, I don't try it in fluorescence, but i think that must work too (the signal with DAB is very intense, and even thin process are stained)
Good luck  :o
ossy
jossy@neurobio.pitt.edu

Offline mhirano

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Thanks
« Reply #4 on: July 23, 2004, 12:15:37 PM »
Thank you, Jossy.
I am not sure that my supervisor allows me to purchase a new antibody, but I shall try.

Offline mhirano

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About anti-GFAP Ab
« Reply #5 on: August 02, 2004, 06:08:30 PM »
Dear Jossy,

I should like to order DAKO's rabbit anti-GFAP polyclonal Ab based on your advice. My supervisor agreed with me, but providing that:
1. You have stained mouse sections (not human sections).
2. Your sections were formalin-fixed, paraffin-embedded sections (not frozen sections).

Would you mind making these points clear?
I am looking forward to your reply.

Sincerely

Offline immunoem

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Need glia marker
« Reply #6 on: August 03, 2004, 03:06:06 AM »
Hi,
anti-GFAP from Biogenex (http://www.biogenex.net/) works fine on mouse wax sections fixed with 10% formalin or 4% pfa (no need for PIER or HIER). As Abs go, it's not too expensive either. Still looking for an equivalent microglia marker. Any suggestions?
Cheers.
Ian

Offline Jossy

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Need glia marker
« Reply #7 on: August 03, 2004, 10:38:50 AM »
Sorry, but I worked with rat brain slices (cut in criostate) , fixed with PFA 4%, I don't know if work in formalin-fixed, paraffin-embedded sections. you should check that.
A marker for microglia (used in the same conditions that GFAP Ab, but in the data sheet say that work in formalin-fixed, paraffin-embedded sections) is OX-42 from Chemicon.
here is data sheet from OX-42 http://www.abcam.com/?datasheet=1211

good luck  :D
ossy
jossy@neurobio.pitt.edu

Offline mhirano

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Need glia marker
« Reply #8 on: August 04, 2004, 12:55:46 PM »
Thank you, Ian and Jossy.
Anti-GFAP from Biogenex seems to be the more suitable for my experiments than Ab from DAKO. And of course, the cheaper the better.

Need glia marker
« Reply #8 on: August 04, 2004, 12:55:46 PM »