FOXP1 (JC12) IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates nuclear staining of large CD20-positive malignant cells.
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The ultraView Universal DAB Detection Kit is specifically designed for performing immunohistochemistry (IHC) on the Ventana® NexES and BenchMark automated systems. The kit detects mouse IgG and IgM and rabbit IgG primary antibodies whereby the specific antibody is located by a cocktail of enzyme-labeled secondary antibodies. This complex is then visualized with hydrogen peroxidase substrate and DAB chromogen, producing a brown precipitate. The ultraView Universal DAB Detection Kit incorporates Ventana-unique Multimer technology, whereby the HRP enzyme is directly conjugated to the secondary antibody. This smaller molecule improves sensitivity and moreover, the biotin-free chemistry minimizes background staining by eliminating staining of endogenous biotin. The kit contains 5 pre-diluted and ready to use dispensers comprising DAB Inhibitor, HRP Multimer, DAB Chromogen, DAB H2O2, Copper and provides enough for 250 tests
I am using the kit for IHC staining of slides prepared from formalin fixed paraffin embedded tissue (FFPET) on the BenchMark Classic Automated system (from Ventana). I have used the kit in conjunction with a variety of primary antibodies either supplied in pre-diluted dispensers or as concentrated antibodies which I manually titrate.
4 micron thin sections of FFPET mounted on SuperFrost Plus slides.
Sections from FFPET biopsies are cut using the microtome at 4 micron thickness and mounted onto SuperFrost Plus slides. For each slide, Ventana labels are then printed which include a unique barcode containing all protocol information. The slides are then loaded onto the BenchMark carousel and read by the Ventana Barcode Reader. The automated Benchmark system puts the slides through a series of user-defined de-paraffinization and antigen retrieval steps before commencing with the antibody staining. The primary antibody is either applied automatically if in a pre-diluted dispenser or otherwise manually titrated onto the slide. The pre-diluted dispensors of the ultraView Universal DAB Detection Kit then provide all reagents required for staining. The counterstain and post-counterstain comprise Haematoxylin (cat #760-2021) and bluing reagent (cat #760-2037) and are purchased separately.
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Low levels of target antigen were detected with specific staining and showing crisp cell morphology.
As the ultraView Universal DAB Detection Kit contains HRP Multimer technology there is minimal background staining due to endogenous biotin exposed during antigen unmasking. This can result in crisp and sensitive staining, reduced cell conditioning and primary antibody incubation times and eliminate the need for blocking reagents.