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"Value": "Upon receipt and when not in use, store at 2-8°C. Do not freeze. This detection kit can be used immediately after removal from the refrigerator.",
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"Value": "VENTANA Red ISH DIG Detection Kit contains sufficient reagent for 60 tests.
- One 6 mL dispenser VENTANA Red ISH DIG NP reagent contains an anti-DIG hapten-labeled antibody (~7.5 μg/mL) in a protein and phosphate containing buffer with 0.05% ProClin 300 solution, a preservative.
- One 6 mL dispenser VENTANA Red ISH DIG NP AP reagent contains an anti-NP alkaline phosphatase (AP) enzyme conjugate (~10 μg/mL) solution in a protein containing buffer including 0.10% ProClin 300 solution, a preservative.
- One 12 mL dispenser VENTANA Red ISH DIG pH Enhancer reagent contains < 3% MgCl2 w/v in a Tris buffer solution including 0.05% ProClin 300 solution, a preservative.
- One 6 mL dispenser VENTANA Red ISH DIG Naphthol reagent contains < 8 g/L naphthol in a Tris buffer solution including 0.05% ProClin 300 solution, a preservative.
- One 12 mL dispenser VENTANA Red ISH DIG Fast Red Chromogen reagent contains < 3 g/L Fast Red KL salt in acetate buffer solution including 0.05% ProClin 300 solution, a preservative.
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"Name": "Background Information",
"Value": "In general, in situ hybridization (ISH) uses labeled probes to detect specific DNA or RNA target sequences in fixed tissue sections. Target sequences are exposed by heating the tissue and probe solution to denature nucleic acids. The reaction is then cooled allowing the labeled nucleic acid probe to hybridize to its complementary nucleic acid sequence in the tissue.
The hybridization of the probe to the nucleic acid sequence is visualized with an indirect detection method. The most common indirect techniques use a secondary antibody directed against the hapten of primary antibody (anti-hapten) and an enzyme linked with a corresponding substrate-chromogen system. This combination results in a colored precipitate at the site of specific probe binding. The VENTANA Red ISH DIG Detection Kit uses the indirect method to visualize complementary nucleic acid sequences by depositing a red colored precipitate. ",
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"Value": "The VENTANA Red ISH DIG Detection Kit is an indirect system for detecting DIG-labeled targets. The kit is intended to identify targets by chromogenic red in situ hybridization (ISH) in sections of formalin-fixed, paraffin-embedded tissue that are stained on BenchMark IHC/ISH instruments.
This product should be interpreted by a qualified reader in conjunction with histological examination, relevant clinical information, and proper controls.",
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"Value": "The VENTANA Red ISH DIG Detection Kit detects probes labeled with digoxigenin (DIG) bound to a sequence in formalin-fixed paraffin-embedded (FFPE) tissue sections. This detection kit includes the following dispensers: mouse anti-DIG primary antibody labeled with nitropyrazole (NP), mouse anti-NP secondary antibody conjugated to Alkaline Phosphatase (AP), pH Enhancer, Naphthol, and Fast Red. Following development of the SISH signal, the slide is incubated with the NP-labeled mouse anti-DIG primary antibody, which binds to the DIG hapten on the probe. The anti-hapten primary antibody is detected with the mouse anti-NP conjugated to AP enzyme. The slide is incubated with the pH Enhancer solution, which provides the proper salt components and concentrations and buffered pH for optimal AP enzyme performance. Next, naphthol phosphate is applied, which serves as the substrate for the AP enzyme (AP dephosphorylates naphthol). Fast Red, added to the slide next, combines with the dephosphorylated naphthol to form a red precipitate, which is readily visualized by light microscopy. Figure 1 illustrates the Red ISH DIG reaction. The specimen is then counterstained with Hematoxylin II for interpretation by light microscopy.
The specific probe is located by a hapten-specific antibody, then by an enzyme-labeled secondary antibody. The complex is then visualized with naphthol substrate and Fast Red chromogen, which produces a red precipitate that is readily detected by light microscopy. The staining protocol consists of numerous steps in which reagents are incubated for pre- determined times at specific temperatures. At the end of each incubation step, the BenchMark IHC/ISH instrument washes the sections to remove unbound material and applies a liquid coverslip which minimizes the evaporation of the aqueous reagents from the slide. Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes, which may or may not be associated with positive staining for the probe.
For more detailed information on instrument operation, refer to the appropriate User Guide.Figure 1 illustrates the indirect detection",
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"Language": "en",
"Country": "XG",
"Code": "Product Purpose"
}
]
}
}
]
}
