Abcam Ad

Author Topic: EtOH fixed paraffin section (Ag-retrieval)  (Read 4931 times)

0 Members and 1 Guest are viewing this topic.

Offline tomtomciao

  • Newbie
  • *
  • Posts: 2
EtOH fixed paraffin section (Ag-retrieval)
« on: July 04, 2005, 10:24:41 AM »
Hi all,

Although the antigen retrieval technique is basically for the formaline fixed paraffin section, have somebody tested the technique with Ethanol fixed paraffin embedded sections? Will it help for the staining intensity? If so, what should I care for the condition?

thanks for your help

tom

EtOH fixed paraffin section (Ag-retrieval)
« on: July 04, 2005, 10:24:41 AM »

Offline ole

  • SilverMember
  • *****
  • Posts: 88
EtOH fixed paraffin section (Ag-retrieval)
« Reply #1 on: July 05, 2005, 04:28:31 PM »
Hi Tom

You simply have to try for yourself. I am not very experienced in this (and i dont think many is). In my experience there is usually no need to do retrieval on ethanol fixed paraffine embedded tissue (EFPET). The "few" antibodies i stain on the two different processed tissues, stains about the same in FFPET (with retrieval) and in EFPET (without retrieval). In fact some cytokeratin markers stain stronger in EFPET (without retrieval) than in FFPET (with retrieval), but thats due to the different properties for the two different fixatives.
Some nuclear antigens are difficult to stain in EFPET and in such cases HIER might enhance the staining. Epitope retrieval easily ruins the morhology in EFPET and i have had not much luck using temp >100deg/C - poor morphology. You might enhance the staining of certain nuclear antigens using a waterbath and temperatures around 95 deg/C.

I have not tried more than about 20 antibodies on EFPET so my experience are limited.

Ole

EtOH fixed paraffin section (Ag-retrieval)
« Reply #1 on: July 05, 2005, 04:28:31 PM »