Permanent HRP Green, 1 kit (100 ml)

Intended use: Permanent HRP Green Kit is intended for immunohistochemical and in situ-hybridisation staining procedures with horse radish peroxidase (HRP). It results in the formation of a green precipitate at the location of the target antigen or target nucleic acid. The precipitate is insoluble in organic solvents and can be observed by light microscopy. The HRP Green precipitate shows a very good contrast to red chromogenic substrates used with alkaline phosphatase detection systems and is therefore especially recommended for double stains. Permanent HRP Green Kit is intended for research use only.

Reagents provided: 100 ml HRP Green Substrate Buffer 3 ml HRP Green Chromogen 1 Dilution Vial

Storage and handling: The solutions should be stored at 2-8°C without fur ther dilution. Please store the reagents in a dark place and do not freeze them. Under these conditions the solutions are stable up to the expiry date indicated on the label. Do not use product after the expiry date. The working solution should be prepared freshly at the day of use. It is stable for at least 4 hours. Excess working solution should be disposed. A positive and a negative control have to be carried out in parallel to the test material. If you observe unusual staining or other deviations from the expected results which could possibly be caused by the kit reagents please contact our technical support.

Reagent preparation: 1) Pipette 1 ml HRP Green Substrate Buffer into the provided dilution vial. 2) Add 1 drop (30 µl) of HRP Green Chromogen. Mix thoroughly. 3) The resulting working solution is stable for at least 4 hours. If you want to prepare other quantities of the working solution, please use the same ratio HRP Green Substrate and HRP Chromogen.

Procedure: 1) Rinse the slide with wash buffer after the previous incubation step. 2) Apply the Permanent HRP Green working solution onto the slide. Incubate for 2-10 minutes. 3) Rinse with distilled H2O. 4) Counterstain with haematoxylin for about 30 seconds up to 5 minutes (depending on the desired staining intensity). 5) Rinse with distilled H2O. 6) Blueing in tap water for 5 minutes. 7) Dehydrate through a graded series of ethanol and clear in xylene. Do not exceed incubation times of 30 sec per dehydration step. Use only high purity xylene. Mount with a permanent mounting medium. Note: The colour intensity can be intensified by increasing the chromogen concentration (up to 3 drops or 90 µl chromogen) in the working solution. Lithium carbonate may have a negative effect on the staining result. We recommend to only bluing in tap water. Occasionally precipitates may appear in the HRP Green Chromogen solution. This doesn’t affect the staining result.

Expected results: During the reaction of the substrate with horse radish peroxidase in presence of the Permanent HRP Green chromogen, a green precipitate is formed at the location of the target antigen or nucleic acid. The precipitate is insoluble in organic solvents and can be observed by light microscopy.

References: Elias JM Immunohistopathology – A practical Approach to Diagnosis ASCP Press 2003/Nadji M and Morales AR Ann N.Y. Acad Sci 420:134-139, 1983

• SKU: SANMON-APP210
• Manufacturer: Sanbio / Monosan
• Manufacturer SKU: MON-APP210
• Package Unit: 1 kit
• Quantity Unit: PAK
• Application: Immunofluorescence, Immunohistochemistry (frozen), Immunohistochemistry (paraffin)