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Monosan Pronase
Monosan Pronase
Artikelnummer
SANMON-APP154
Verpackungseinheit
250 ml
Hersteller
Sanbio / Monosan
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Intended use: Pronase Solution is a ready-to-use solution developed for enzymatic epitope retrieval on formalin-fixed tissue sections on slides. This procedure (sometimes called PIER, Protease Induced Epitope Retrieval) is primarily used in immunohistochemical staining procedures. Proteolytic pre-treatments are also used in protocols for in situhybridization. Pronase solution is intended for research use only.
Summary and explanation: Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Enzymatic digestion with proteolytic enzymes (PIER) restores structures of the epitopes making them more accessible to specific antibodies. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Reagents provided: 250 ml Pronase Solution (Ready-To-Use)
Storage and handling: The solution should be stored in aliquots at -20°C without further dilution. The solution should be aliquoted in order to avoid repeated freeze and thawing. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date.
Principle of method: Pronase Solution is a ready-to-use solution for enzymatic epitope retrieval.
Reagent preparation: Pronase solution is ready-to-use and should be at room temperature prior to use.
Procedure: Pronase Solution is suitable for enzymatic epitope retrieval carried out after the dewaxing and rehydration of the tissue sections. 1. Cover deparaffinised and rehydrated tissue sections with ready-to-use Pronase Solution. 2. Incubate for 15 - 20 minutes at room temperature. The optimal incubation time needs to be elaborated by the operator. 3. Rinse carefully (3 x), first with distilled water followed by buffer. 4. Proceed with immunohistological staining as usual.
Expected results: During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
Summary and explanation: Immunohistochemical staining procedures consist of sequential incubation steps with blocking solutions, antibodies and secondary reagents, enzymes and chromogenic substrates carried out on tissue sections. These tissue sections are mostly prepared out of formalin-fixed paraffin-embedded tissue blocks. Cellular structures are very effectively stabilised by formalin fixation which results in optimal morphological preservation of the sample. On the other hand the formalin fixation leads to strong cross-links between proteins. This means that epitopes of antigens are being masked and often are no longer accessible for primary antibodies. In order to enable primary antibodies to bind to antigens the epitopes have to be recovered. Enzymatic digestion with proteolytic enzymes (PIER) restores structures of the epitopes making them more accessible to specific antibodies. Heat induced epitope retrieval (HIER) in buffer solutions of different compositions and pH-values is another way of recovering epitopes. The primary antibody used determines the appropriate method.
Reagents provided: 250 ml Pronase Solution (Ready-To-Use)
Storage and handling: The solution should be stored in aliquots at -20°C without further dilution. The solution should be aliquoted in order to avoid repeated freeze and thawing. Under these conditions the solution is stable up to the expiry date indicated on the label. Do not use product after the expiry date.
Principle of method: Pronase Solution is a ready-to-use solution for enzymatic epitope retrieval.
Reagent preparation: Pronase solution is ready-to-use and should be at room temperature prior to use.
Procedure: Pronase Solution is suitable for enzymatic epitope retrieval carried out after the dewaxing and rehydration of the tissue sections. 1. Cover deparaffinised and rehydrated tissue sections with ready-to-use Pronase Solution. 2. Incubate for 15 - 20 minutes at room temperature. The optimal incubation time needs to be elaborated by the operator. 3. Rinse carefully (3 x), first with distilled water followed by buffer. 4. Proceed with immunohistological staining as usual.
Expected results: During the reaction of the substrate with horse radish peroxidase or alkaline phosphatase in the presence of a chromogen, a coloured precipitate is formed at the location of the bound primary antibody. This reaction only takes place if the target antigen is existent in the tissue. The chromogen used determines the colour of the precipitate. The analysis is carried out using a light microscope.
| Artikelnummer | SANMON-APP154 |
|---|---|
| Hersteller | Sanbio / Monosan |
| Hersteller Artikelnummer | MON-APP154 |
| Verpackungseinheit | 250 ml |
| Mengeneinheit | STK |
| Methode | Immunofluorescence, Immunohistochemistry (frozen), Immunohistochemistry (paraffin) |
| Produktinformation (PDF) | Download |
| MSDS (PDF) |
|
Immer für Sie da
Gernot Ensinger, M.Sc.
Laurent Haze
