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ultraView Red ISH DIG Detection Kit detects digoxigenin (DIG) labeled probes bound to target sequences using red color in situ hybridization (Red ISH) in paraffin embedded tissue sections. First, the section is hybridized with a DIG-labeled probe, followed by incubation with a mouse monoclonal anti-DIG antibody, which binds to the DIG hapten on the probe. A Multimer solution, a goat anti-mouse IgG with an alkaline phosphatase (AP) enzyme, is applied to detect the mouse anti-DIG antibody. The section is incubated with the pH Enhancer solution, which primes the AP enzyme and provides the proper salt components/concentrations and buffered pH for optimal AP performance. The section is incubated with Naphthol, which provides the substrate for the AP enzyme (AP dephosphorylates Naphthol). Fast Red is added to the section, which combines with Naphthol to produce a red precipitate, which is readily visualized by light microscopy.
Figure 1 illustrates the Red ISH reaction. The specimen is then counterstained with Hematoxylin II for interpretation by light microscopy.
Figure 1. Red ISH Reaction for DIG-labeled Probe Detection
The
ultraView Red ISH DIG Detection Kit is optimized for use with VENTANA probes, accessory reagents and VENTANA BenchMark XT or BenchMark ULTRA automated slide stainers. The staining protocol consists of numerous steps in which reagents are incubated for precise times at specific temperatures. At the end of each incubation step, the automated slide stainer washes the sections to remove unbound material and applies a liquid coverslip, which minimizes evaporation of aqueous reagents from the slide.
1 Results are interpreted using a light microscope and aid in the differential diagnosis of pathophysiological processes.
For more detailed information on instrument operation, refer to the appropriate VENTANA automated slide stainer Operator's Manual.
1. Elias JM, Gown AM, Nakamura RM, Wilbur DC, Herman GE, Jaffe ES, Battifora H, Brigati DJ. Quality control in immunohistochemistry. Report of a workshop sponsored by the Biological Stain Commission. Am J Clin Pathol. 1989;92(6):836- 843.",
"Language": "en",
"Country": "XG",
"Code": "Principle"
},
{
"Name": "Content",
"Value": "
ultraView Red ISH DIG Detection Kit provides reagents sufficient for 100 tests.
- One 10 mL dispenser ultraView Red ISH DIG contains < 5µg/mL mouse monoclonal anti-DIG antibody in a buffer.
- One 10 mL dispenser ultraView Red ISH DIG Multimer contains < 50µg/mL goat anti-mouse IgG alkaline phosphatase (AP) conjugate in a buffer with 0.05% ProClin 300, a preservative.*
- One 20 mL dispenser ultraView Red ISH DIG pH Enhancer contains ≤3%(v/v) MgCl2 in a Tris buffer with ProClin 300, a preservative
- One 10 mL dispenser ultraView Red ISH DIG Naphthol contains ≤1.0%(w/v) Naphthol in Tris buffer with ProClin 300, a preservative.
- One 20 mL dispenser ultraView Red ISH DIG Fast Red contains ≤1.0%(w/v) Fast Red KL Salt in acetate buffer with ProClin 300, a preservative.
*For preservative information please refer to the Warnings and Precautions section of the method sheet.",
"Language": "en",
"Country": "XG",
"Code": "Content"
},
{
"Name": "Product Purpose",
"Value": "Ventana Medical Systems, Inc’s (Ventana)
ultraView Red ISH DIG Detection Kit is an indirect, biotin-free system for detecting DIG labeled probes. 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 VENTANA BenchMark XT automated slide stainer or VENTANA BenchMark ULTRA automated slide stainer.
The clinical interpretation of any staining, or the absence of staining, must be complemented by histological examination and evaluation of proper controls. Evaluation must be made by a qualified reader within the context of the patient’s clinical history and other diagnostic tests.
This detection kit is intended for
in vitro diagnostic (IVD) use.",
"Language": "en",
"Country": "XG",
"Code": "Product Purpose"
}
]
}
}
]
}