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Electron Microscopy Sciences

Technical Data Sheets

Retriever FAQ

EMS Catalog #62706

What tissue slides should I use for sections processed in Retriever?

Usually Superfrost+ performs rather well. To ensure safety of your most valuable material (especially tissue arrays) we suggest using always activated slides. You can easily prepare them in your lab following the instructions in the Manual.

I used in Retriever the EDTA pH8.0 buffer and the tissues detached from the slides

This is a general problem with compatibility of tissue slides with EDTA buffer. If you have used just regular Superfrost slides, and sometimes even Superfrost + tissue may detach in the buffer. We recommend using Hybond or Superfrost Gold slides.

The tissue did not detach from the slide, but after Retriever some of the tissue areas lost morphology

Sometimes these are internal areas of the not sufficiently fixed, or improperly processed (paraffin embedding) tissues. In principle these tissues may be used. We advice to add to your buffer glycerol to 5% or 10% and re-adjust again the pH to the required for this type of buffer. This would make processing of the tissue less stringent.

I am used to Tris Buffer for antigen unmasking, would it work in Retriever?

We do not recommend using Tris buffer with Retriever.

Can I use in Retriever standard citrate (pH 6.0) and EDTA (pH 8.0) buffers

Yes, you can.

What if tissues are fixed with other than formalin fixatives? Can they be processed in Retriever?

One of the frequently used fixatives is formalin/acetic acid fixative. Laboratories in France and Belgium, where it is frequently used in routine pathology, report that Retriever works well with these tissues, and even superior to other means of antigen unmasking when it comes to some antigens, estrogen receptor in particular.

Do all antibodies work in IHC after Retriever?

Most of them do. If antibody works well in immunoblotting, it, as a rule works in IHC on Retriever -processed tissue. When you have a choice, look for antibodies prepared against a fusion protein (fragment) that work in blotting. For the majority of these antibodies start with buffer A or citrate buffer (pH 6.0). Usually they give no problems. Antibodies prepared against peptide epitope working in immunoblotting, also work on sections as a rule, unless the peptide epitope contains too many Lys; this may lead to epitope inactivation, an irreversible one. Use buffer A or buffer U. Most of the problems are with monoclonal antibodies prepared against native epitopes and mainly used in flow cytometry. These often require testing a variety of buffers to find an appropriate one.

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