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Author Topic: DAB GFAP staining on Parafin Sections  (Read 3578 times)

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Offline ADoering

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DAB GFAP staining on Parafin Sections
« on: May 01, 2014, 01:42:05 PM »
Hello Group,

I am struggeling with getting my GFAP staining to work. I use paraffin embedded tissue and it should not be a problem to  get a staining. I am quite expreienced and at a loss here (changed AB, AB conc., HIAR, ect)/ Any suggestions?

Thanks

DAB GFAP staining on Parafin Sections
« on: May 01, 2014, 01:42:05 PM »

Offline gula

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Re: DAB GFAP staining on Parafin Sections
« Reply #1 on: May 01, 2014, 02:32:25 PM »
Just a bad antibody? Did you test antibodies from different suppliers?
It's a rare but existing phenomen. ;-)
gula

Offline ADoering

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Re: DAB GFAP staining on Parafin Sections
« Reply #2 on: May 01, 2014, 05:09:47 PM »
Thanks, tried 2 different AB, using now the poly rb from Dako in 1:500 as recommended. HIAR was done in a steamer for 40 min Citrate Buffer...It drives me insane as I am very experienced in IHC and this is normally such an easy stain....


Offline luisa

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Re: DAB GFAP staining on Parafin Sections
« Reply #3 on: May 02, 2014, 03:41:59 AM »
For me, polyclonal anti-GFAP from Dako works great using enzymatic retrieval with PK at 20 ug/ml, 39ēC, 30 min, at 1:1000.
Luisa

Offline luisa

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Re: DAB GFAP staining on Parafin Sections
« Reply #4 on: May 02, 2014, 03:42:35 AM »
Sorry, 37ēC...
Luisa

Offline gula

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Re: DAB GFAP staining on Parafin Sections
« Reply #5 on: May 02, 2014, 12:31:13 PM »
Which detection system do you use? IHC-platform?
Have your tried several specimens? perhaps a matter of pretreatment, fixation...
Do you dry the sections prior to staining? some antigens are sensitive to too hot/long dry heat.
gula

Re: DAB GFAP staining on Parafin Sections
« Reply #5 on: May 02, 2014, 12:31:13 PM »